Optimized peptides for targeting human nerves and their use in image guided surgery, diagnostics and therapeutic delivery

ABSTRACT

The present invention provides methods for guiding preservation of human neurons or human nerves during surgery by administering a fluorescently-labeled peptide that specifically binds to the human neurons or human nerves. The invention further provides human neuron or nerve targeting molecules comprising fluorescently-labeled peptides that specifically bind to human neurons or human nerves and compositions thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.16/780,782, filed Feb. 3, 2020, which is a continuation of U.S.application Ser. No. 16/635,133, filed Jan. 29, 2020, which is a 371National Phase of PCT/US2018/045054, filed Aug. 2, 2018, which claimspriority to U.S. Provisional Application No. 62/659,612, filed Apr. 18,2018, and U.S. Provisional Application No. 62/540,510, filed Aug. 2,2017, all of which are incorporated herein by reference in theirentireties.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH OR DEVELOPMENT

This invention was made with Government support under EB008122 andEB014929 awarded by the National Institutes of Health. The Governmenthas certain rights in the invention

BACKGROUND OF THE INVENTION

Preservation of human neurons and human nerves is one of the mostimportant goals of any surgical procedure, because accidentaltransection of neuron or nerves leads to significant morbidity. Nervesare typically identified by their elongated whitish appearance andrelationship to nearby structures or by electrophysiological studies.However, in instances such as trauma, tumor involvement, inflammation,or infection, nerve identification using these criteria can bedifficult. Therefore, there is a need for methods of reliably andconclusively identifying neuron or nerves which overcome thedeficiencies in the art.

Neuron or nerve identification prior to direct exposure during surgeryor confirmation of neuron or nerve identity in instances of uncertaintyfollowing direct exposure is accomplished by electromyographic (EMG)monitoring. This technique, however, has the disadvantage of notproviding visual feedback to the operating surgeon. Thus, even if anerve has been identified in one location, either through accidental orpurposeful stimulation, there is no visual guidance to the operatingsurgeon as to how far away from the stimulation site the nerve lies orthe direction of travel the nerve takes away from the stimulation site.Furthermore, EMG only traces motor pathways, not sensory fibers. EMGfails if neuron or nerve conduction or neuromuscular transmission istemporarily blocked anywhere distal to the recording site. Such blockadeeasily occurs due to neuron or nerve compression, trauma, localanesthetics, or neuromuscular blockers.

Neuron or nerve labeling primarily depend on retrograde or anterogradetracing of individually identified axonal tracts via the use offluorescent dyes. However, methods of labeling neuron or nerves bylocally applied fluorescent tracers have several disadvantages. First,this technique can label only one neuron or nerve fiber tract at a time,depending on where the dye has been injected. Second, this techniqueresults in only limited labeling of fluorescent dyes along the axonaltracts, because retrograde axonal tracers typically accumulate in theneural cell body. Third, retrograde transport is relatively slow (on theorder of millimeters per day) and therefore takes a long time to labelhuman neuron or nerves, which are often longer than a meter, such as inthe case of the sciatic neuron or nerve and its arborizations. Fourth,the application of fluorescent dyes to innervation targets such asdirect intramuscular injections to label motor neuron or nerves istypically messy with a variable amount of the tracer dye remaining atthe injection site. As dissection of neuron or nerves depends onaccurate visualization of adjacent structures prior to encounteringthem, a surgical site that is contaminated with fluorescent dyes wouldnot be desirable. Finally, the direct injection of the fluorescent dyeitself may be damaging to the target organs or neuron or nerve ofinterest, either by mechanical damage or by the very high localconcentration of dye and vehicle at the injection site.

There has been a need in the art to identify peptides capable of bindingto human nerves and neurons, in order to facilitate surgical proceduresand human nerve protection.

Nerve-homing peptides sequences were previously identified by theirability to bind mouse nerves for laboratory research. However, thepeptide sequences described in the present application were identifiedby their ability to bind human nerves, following systemic intravenousinjection into human patients and as such these peptides meet the needof being able to more specifically and effectively bind to human nervescompared to previous sequences. The present invention provides peptidesequences that selective bind to human nerves and/or neurons, as well asmethods of using those sequences in surgical procedures, for example topreserve nerves and/or to avoid nerve damage during such procedures.

BRIEF SUMMARY OF THE INVENTION

Disclosed herein, in certain embodiments, are targeting moleculescomprising a peptide that specifically binds to a human neuron, humannerve, or component of either. In some embodiments, the peptide isselected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16),Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4),Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSS (HNP401-N-2with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22);EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS (HNP401-N-8; SEQ IDNO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV(HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27),SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 withGG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120),SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121),SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124),

5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104),PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118),EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG(HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG(HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GGlinker; SEQ ID NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ IDNO:124), and/or combinations thereof.

In some embodiments, the human neuron or nerve targeting molecule thatspecifically binds to a human neuron or nerve, or component of either,wherein said targeting molecule comprises a peptide selected from thegroup consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104), PWEEPYYVVKKSSGG (HNP401-N-4with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GGlinker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ IDNO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121),SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and/or combinations thereof.

In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting: of SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3).

In some embodiments, the targeting molecule comprises a peptide selectedfrom the group consisting of SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21), and SGQVPWEEPYYVVKK(HNP401-C-2; SEQ ID NO:25).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1).

In some embodiments, the targeting molecule comprises a peptideWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2).

In some embodiments, the targeting molecule comprises the peptideDLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

In some embodiments, the targeting molecule comprises the peptideAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

In some embodiments, the targeting molecule comprises the peptideAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

In some embodiments, the targeting molecule comprises the peptideAc-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

In some embodiments, the targeting molecule comprises the peptideAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

In some embodiments, the targeting molecule comprises the peptideAc-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9).

In some embodiments, the targeting molecule comprises the peptideAc-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14).

In some embodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).

In some embodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

In some embodiments, the targeting molecule comprises the peptidePWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22).

In some embodiments, the targeting molecule comprises the peptideEEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23).

In some embodiments, the targeting molecule comprises the peptidePYYVVKKSS (HNP401-N-8; SEQ ID NO:24).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

In some embodiments, the targeting molecule comprises the SGQVPWEEPYYVV(HNP401-C-4; SEQ ID NO:26).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEP (HNP401-C-8; SEQ ID NO:28).

In some embodiments, the targeting molecule comprises the peptidePWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118).

In some embodiments, the targeting molecule comprises the peptideEEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119).

In some embodiments, the targeting molecule comprises the peptidePYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

In some embodiments, the targeting molecule comprises the peptide5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a cargo. In some embodiments, the cargo is a drug, afluorescent moiety, a photosensitizing agent, or a combination thereof.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a drug.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a drug selected from the group consisting of: anantihistamine, a GABA receptor modulator, a neurotransmitter reuptakeinhibitor, a local anesthetic, an anticholinergic, a sodium channelblocker, a calcium channel blocker, a thyrotropin-releasing hormone, aγ-secretase inhibitor, an AMPA receptor agonist or antagonist, an NMDAreceptor agonist or antagonist, an mGlu receptor agonist or antagonist,a growth factor, an antiemetic agent, a corticosteroid; a cytotoxicagent; an antioxidant, an iron chelator, a mitochondrial modulator, asirtuin modulator, a nitric oxide (NO) and/or nitric oxide synthase(NOS) modulator, a potassium channel agonist or antagonist, a purigenicreceptor agonist or antagonist, and/or combinations thereof.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a drug selected from the group consisting of:benzocaine; carticaine; cinchocaine; cyclomethycaine; lidocaine;prilocaine; propxycaine; proparacaine; tetracaine; tocainide; andtrimecaine; methotrexate; cyclophosphamide; thalidomide; paclitaxel;pemetrexed; pentostatin; pipobroman; pixantrone; plicamycin; platonin;procarbazine; raltitrexed; rebeccamycin; rubitecan; SN-38;salinosporamide A; satraplatin; streptozotocin; swainsonine; tariquidar;taxane; tegafur-uracil; temozolomide; testolactone; thioTEPA;tioguanine; topotecan; trabectedin; tretinoin; triplatin tetranitrate;tris(2-chloroethyl)amine; troxacitabine; uracil mustard; valrubicin;vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar;carbamazepine; oxcarbazepine; phenytein; valproic acid; sodiumvalproate; cinnarizine; flunarizine; nimodipine; brain-derivedneurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glialcell-line derived neurotrophic factor (GDNF); neurotrophin-3;neurotrophin-4; fibroblast growth factor (FGF) receptor; insulin-likegrowth factor (IGF); and/or combinations thereof.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a fluorescent moiety.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a fluorescent moiety selected from the groupconsisting of: a fluorescent protein, a fluorescent peptide, afluorescent dye, and/or combinations thereof.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a fluorescent moiety selected from the groupconsisting: of a xanthene; a bimane; a coumarin; an aromatic amines; abenzofuran; a fluorescent cyanine; a carbazole; a dicyanomethylenepyrane; polymethine; oxabenzanthrane; pyrylium; carbostyl; perylene;acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene;pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metalchelate complexes; rare-earth metal chelate complexes; derivativesthereof, and/or combinations thereof.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a fluorescent moiety selected from the groupconsisting: of 5-carboxyfluorescein; fluorescein-5-isothiocyanate;6-carboxyfluorescein; 5(6)-carboxyfluorescein;tetramethylrhodamine-6-isothiocyanate; 5-carboxytetramethylrhodamine;5-carboxy rhodol derivatives; tetramethyl and tetraethyl rhodamine;diphenyldimethyl and diphenyldiethyl rhodamine; dinaphthyl rhodamine;rhodamine 101 sulfonyl chloride; Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy 7,indocyanine green, IR800CW, cyan fluorescent protein (CFP), EGFP, 6-FAM,FAM, fluorescein, 5,6-dicarboxyfluorescein, 5-(and 6)-sulfofluorescein,sulfonefluorescein, succinyl fluorescein, 5-(and 6)-carboxy SNARF-1,carboxyfluorescein sulfonate, carboxyfluorescein zwitterion,carboxyfluorescein quaternary ammonium, carboxyfluorescein phosphonate,carboxyfluorescein GABA, carboxyfluorescein-cys-Cy5,5′(6′)-carboxyfluorescein, fluorescein glutathione, and/or combinationsthereof.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a photosensitizing agent.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a photosensitizing agent selected from the groupconsisting of: a porphyrin, chlorin, and dye.

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a photosensitizing agent selected from the groupconsisting of: porphyrin, protoporfin IX, purlytin, verteporfin, HPPH,temoporfin, methylene blue, photofrin, protofrin, hematoporphyrin,Talaporfin, benzopophyrin derivative monoacid, 5-aminileuvolinic acid,Lutetium texaphyrin, metallophthalocyanine,metallo-naphthocyaninesulfobenzo-porphyrazine,metallo-naphthalocyanines\, zinc tetrasulfophthalocyanine,bacteriochlorins, metallochlorins, chlorine derivative,Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein(DBF), IR700DX, naphthalocyanine, porphyrin derivative, and/orcombinations thereof.

In some embodiments, provided is a method of identifying a human neuronor nerve comprising contacting the human neuron or nerve with atargeting molecule comprising (a) a peptide that specifically binds tothe human neuron or nerve, or component of either, and (b) a fluorescentmoiety, wherein said targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104), and/or combinations thereof.

In some embodiments, the human neuron or nerve targeting moleculecomprises a peptide selected from the group consisting of:SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2), and DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

In some embodiments, the human neuron or nerve targeting moleculecomprises a peptide selected from the group consisting of:SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC(HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC(HNP401-N-2 with GGC linker; SEQ ID NO:7), QVPWEEPYYVVKKSS (HNP401-N-2;SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ IDNO:21), and SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1).

In some embodiments, the targeting molecule comprises the peptideWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2).

In some embodiments, the targeting molecule comprises the peptideDLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

In some embodiments, the targeting molecule comprises the peptideAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

In some embodiments, the targeting molecule comprises the peptideAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

In some embodiments, the targeting molecule comprises the peptideAc-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

In some embodiments, the targeting molecule comprises the peptideAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

In some embodiments, the targeting molecule comprises the peptideAc-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9).

In some embodiments, the targeting molecule comprises the peptideAc-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14).

In some embodiments, the targeting molecule comprises the peptideDTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16).

In some embodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).

In some embodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

In some embodiments, the targeting molecule comprises the peptidePWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22).

In some embodiments, the targeting molecule comprises the peptideEEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23).

In some embodiments, the targeting molecule comprises the peptidePYYVVKKSS (HNP401-N-8; SEQ ID NO:24).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEP (HNP401-C-8; SEQ ID NO:28).

In some embodiments, the targeting molecule comprises the peptidePWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118).

In some embodiments, the targeting molecule comprises the peptideEEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119).

In some embodiments, the targeting molecule comprises the peptidePYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

In some embodiments, the fluorescent moiety is selected from the groupconsisting of: a fluorescent protein, a fluorescent peptide, afluorescent dye, and/or combinations thereof.

In some embodiments, the targeting molecule comprises the peptide5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

In some embodiments, the fluorescent moiety is selected from the groupconsisting: of a xanthene; a bimane; a coumarin; an aromatic amine; abenzofuran; a fluorescent cyanine; a carbazole; a dicyanomethylenepyrane; polymethine; oxabenzanthrane; pyrylium; carbostyl; perylene;acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene;pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metalchelate complexes; rare-earth metal chelate complexes; derivativesthereof, and/or combinations thereof.

In some embodiments, the fluorescent moiety is selected from the groupconsisting of: 5-carboxyfluorescein (5-FAM);fluorescein-5-isothiocyanate; 6-carboxyfluorescein (6-FAM);5(6)-carboxyfluorescein; tetramethylrhodamine-6-isothiocyanate;5-carboxytetramethylrhodamine; 5-carboxy rhodol derivatives; tetramethyland tetraethyl rhodamine; diphenyldimethyl and diphenyldiethylrhodamine; dinaphthyl rhodamine; rhodamine 101 sulfonyl chloride; Cy3,Cy3B, Cy3.5, Cy5, Cy5.5, Cy 7, indocyanine green, IR800CW, cyanfluorescent protein (CFP), EGFP, 6-FAM, FAM, fluorescein,5,6-dicarboxyfluorescein, 5-(and 6)-sulfofluorescein,sulfonefluorescein, succinyl fluorescein, 5-(and 6)-carboxy SNARF-1,carboxyfluorescein sulfonate, carboxyfluorescein zwitterion,carboxyfluorescein quaternary ammonium, carboxyfluorescein phosphonate,carboxyfluorescein GABA, carboxyfluorescein-cys-Cy5,5′(6′)-carboxyfluorescein, fluorescein glutathione, and/or combinationsthereof.

In some embodiments, provided is a method of delivering a drug to ahuman neuron or nerve comprising contacting the human neuron or nervewith a human neuron or nerve targeting molecule comprising (a) a peptidethat specifically binds to the neuron or nerve, or component of either,and (b) a drug, wherein said targeting molecule comprises a peptideselected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16),Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4),Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124),

5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104),and/or combinations thereof.

In some embodiments, the drug is selected from the group consisting of:an antihistamine, a GABA receptor modulator, a neurotransmitter reuptakeinhibitor, a local anesthetic, an anticholinergic, a sodium channelblocker, a calcium channel blocker, a thyrotropin-releasing hormone, aγ-secretase inhibitor, an AMPA receptor agonist or antagonist, an NMDAreceptor agonist or antagonist, an mGlu receptor agonist or antagonist,a growth factor, an antiemetic agent, a corticosteroid; a cytotoxicagent; an antioxidant, an iron chelator, a mitochondrial modulator, asirtuin modulator, a nitric oxide (NO) and/or nitric oxide synthase(NOS) modulator, a potassium channel agonist or antagonist, a purigenicreceptor agonist or antagonist, and/or combinations thereof.

In some embodiments, the drug is selected from the group consisting of:benzocaine; carticaine; cinchocaine; cyclomethycaine; lidocaine;prilocaine; propxycaine; proparacaine; tetracaine; tocainide; andtrimecaine; methotrexate; cyclophosphamide; thalidomide; paclitaxel;pemetrexed; pentostatin; pipobroman; pixantrone; plicamycin;procarbazine; raltitrexed; rebeccamycin; rubitecan; SN-38;salinosporamide A; satraplatin; streptozotocin; swainsonine; tariquidar;taxane; tegafur-uracil; temozolomide; testolactone; thioTEPA;tioguanine; topotecan; trabectedin; tretinoin; triplatin tetranitrate;tris(2-chloroethyl)amine; troxacitabine; uracil mustard; valrubicin;vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar;carbamazepine; oxcarbazepine; phenytein; valproic acid; sodiumvalproate; cinnarizine; flunarizine; nimodipine; brain-derivedneurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glialcell-line derived neurotrophic factor (GDNF); neurotrophin-3;neurotrophin-4; fibroblast growth factor (FGF) receptor; insulin-likegrowth factor (IGF); and/or combinations thereof.

In some embodiments, provided is a method of delivering aphotosensitizing agent to a human neuron or nerve comprising contactingthe human neuron or nerve with a human neuron or nerve targetingmolecule comprising (a) a peptide that specifically binds to the neuronor nerve, or component of either, and (b) a photosensitizing agent,wherein said targeting molecule comprises a peptide selected from thegroup consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124),

5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104),and/or combinations thereof.

In some embodiments, the method further comprises exposing the humanneuron or nerve to a light source that activates the photosensitizingagent.

In some embodiments, the photosensitizing agent is selected from thegroup consisting of: a porphyrin, chlorin, and dye.

In some embodiments, the photosensitizing agent selected from the groupconsisting of: porphyrin, protoporfin IX, purlytin, verteporfin, HPPH,temoporfin, methylene blue, photofrin, protofrin, hematoporphyrin,Talaporfin, benzopophyrin derivative monoacid, 5-aminileuvolinic acid,Lutetium texaphyrin, metallophthalocyanine,metallo-naphthocyaninesulfobenzo-porphyrazine,metallo-naphthalocyanines\, zinc tetrasulfophthalocyanine,bacteriochlorins, metallochlorins, chlorine derivative,Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein(DBF), IR700DX, naphthalocyanine, porphyrin derivative, and/orcombinations thereof.

In some embodiments, the human neuron or nerve targeting molecule isadministered by systemic intravenous injection a human subject.

In some embodiments, the human neuron or nerve targeting molecule isadministered prior to a surgical procedure. In some embodiments, thesurgical procedure is a cancer surgical procedure. In some embodiments,the surgical procedure is a prostate cancer surgical procedure.

In some embodiments, provided is a pharmaceutical compositioncomprising: (a) a peptide that specifically binds to a human neuron,human nerve, or component of either, and (b) a pharmaceuticallyacceptable excipient, wherein said human neuron or nerve targetingmolecule comprises a peptide selected from the group consisting of:SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104), and/or combinations thereof.

In some embodiments of the composition, the peptide is selected from thegroup consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3).

In some embodiments of the composition, the peptide is selected from thegroup consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20) QVPWEEPYYVVKKSSGG (HNP401-N-2with GG linker; SEQ ID NO:21), and SGQVPWEEPYYVVKK (HNP401-C-2; SEQ IDNO:25).

In some embodiments of the composition, the peptide comprisesSGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1).

In some embodiments of the composition, the peptide comprisesWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2).

In some embodiments of the composition, the peptide comprisesDLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

In some embodiments of the composition, the peptide comprisesAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

In some embodiments of the composition, the peptide comprisesAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

In some embodiments of the composition, the peptide comprisesAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

In some embodiments of the composition, the peptide comprisesAc-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

In some embodiments of the composition, the peptide comprisesAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

In some embodiments of the composition, the peptide comprisesAc-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9).

In some embodiments of the composition, the peptide comprisesAc-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10).

In some embodiments of the composition, the peptide comprisesAc-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

In some embodiments of the composition, the peptide comprisesAc-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

In some embodiments of the composition, the peptide comprisesAc-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).

In some embodiments of the composition, the peptide comprisesAc-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14).

In some embodiments of the composition, the peptide comprisesDTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16).

In some embodiments of the composition, the peptide comprisesQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).

In some embodiments of the composition, the peptide comprisesQVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

In some embodiments of the composition, the peptide comprisesPWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22).

In some embodiments of the composition, the peptide comprisesEEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23).

In some embodiments of the composition, the peptide comprises PYYVVKKSS(HNP401-N-8; SEQ ID NO:24).

In some embodiments of the composition, the peptide comprisesSGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

In some embodiments of the composition, the peptide comprisesSGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26).

In some embodiments of the composition, the peptide comprisesSGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27).

In some embodiments of the composition, the peptide comprises SGQVPWEEP(HNP401-C-8; SEQ ID NO:28).

In some embodiments of the composition, the peptide comprisesPWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118).

In some embodiments of the composition, the peptide comprisesEEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119).

In some embodiments of the composition, the peptide comprisesPYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120).

In some embodiments of the composition, the peptide comprisesSGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121).

In some embodiments of the composition, the peptide comprisesSGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122).

In some embodiments of the composition, the peptide comprisesSGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123).

In some embodiments of the composition, the peptide comprisesSGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

In some embodiments of the composition, the peptide comprises5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

In some embodiments of the composition, the peptide is SGQVPWEEPYYVVKKSS(HNP 401; SEQ ID NO:1).

In some embodiments of the composition, the peptide is WEYHYVDLNWTSQHPQ(HNP 402; SEQ ID NO:2).

In some embodiments of the composition, the peptide is DLPDIIWDFNWETA(HNP 403; SEQ ID NO:3).

In some embodiments of the composition, the peptide isAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

In some embodiments of the composition, the peptide isAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

In some embodiments of the composition, the peptide isAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

In some embodiments of the composition, the peptide isAc-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

In some embodiments of the composition, the peptide isAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

In some embodiments of the composition, the peptide is Ac-EEPYYVVKKSSGGC(HNP401-N-6 with GGC linker; SEQ ID NO:9).

In some embodiments of the composition, the peptide is Ac-PYYVVKKSSGGC(HNP401-N-8 with GGC linker; SEQ ID NO:10).

In some embodiments of the composition, the peptide isAc-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

In some embodiments of the composition, the peptide isAc-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

In some embodiments of the composition, the peptide is Ac-SGQVPWEEPYYGGC(HNP401-C-6 with GGC linker; SEQ ID NO:13).

In some embodiments of the composition, the peptide is Ac-SGQVPWEEPGGC(HNP401-C-8 with GGC linker; SEQ ID NO:14).

In some embodiments of the composition, the peptide is DTHAHAKPRVPAFKSV(HNP 404; SEQ ID NO:16).

In some embodiments of the composition, the peptide is QVPWEEPYYVVKKSS(HNP401-N-2; SEQ ID NO:20).

In some embodiments of the composition, the peptide is QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21).

In some embodiments of the composition, the peptide is PWEEPYYVVKKSS(HNP401-N-4; SEQ ID NO:22).

In some embodiments of the composition, the peptide is EEPYYVVKKSS(HNP401-N-6; SEQ ID NO:23).

In some embodiments of the composition, the peptide is PYYVVKKSS(HNP401-N-8; SEQ ID NO:24).

In some embodiments of the composition, the peptide is SGQVPWEEPYYVVKK(HNP401-C-2; SEQ ID NO:25).

In some embodiments of the composition, the peptide is SGQVPWEEPYYVV(HNP401-C-4; SEQ ID NO:26).

In some embodiments of the composition, the peptide is SGQVPWEEPYY(HNP401-C-6; SEQ ID NO:27).

In some embodiments of the composition, the peptide is SGQVPWEEP(HNP401-C-8; SEQ ID NO:28).

In some embodiments of the composition, the peptide is5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

In some embodiments of the composition, the peptide is PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118).

In some embodiments of the composition, the peptide is EEPYYVVKKSSGG(HNP401-N-6 with GG linker; SEQ ID NO:119).

In some embodiments of the composition, the peptide is PYYVVKKSSGG(HNP401-N-8 with GG linker; SEQ ID NO:120).

In some embodiments of the composition, the peptide is SGQVPWEEPYYVVKKGG(HNP401-C-2; with GG linker; SEQ ID NO:121).

In some embodiments of the composition, the peptide is SGQVPWEEPYYVVGG(HNP401-C-4 with GG linker; SEQ ID NO:122).

In some embodiments of the composition, the peptide is SGQVPWEEPYYGG,(HNP401-C-6 with GG linker; SEQ ID NO:123).

In some embodiments of the composition, the peptide is SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124).

In some embodiments of the composition, the peptide is bound to a cargo.In some embodiments, the cargo is a drug, photosensitizing agent, orfluorescent moiety.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Fluorescence images of exposed sciatic nerves in living wildtype mice following administration of 450 nmols HNP401 in three mice(left) and NP41 (two mice, right). Both left and right sciatic nervesare shown. FAM is the fluorescein attached to the C-terminal lysine ofeach peptide sequence. Images were obtained with a Zeiss Lumar.

FIG. 2: Quantitation of images from FIG. 1 show HNP 401 have similarcontrast (nerve fluorescent intensity/muscle fluorescent intensity) toNP41 (6.2 fold compared to 6.7 fold) with with higher intensity labelingof both nerve and adjacent muscle tissue. 6 nerves, 3 mice total forHNP401 and 2 mice (4 nerves) for NP41.Y axis=average fluorescent (515 nmemission) intensity calculated in imageJ.

FIG. 3: Topical application of Nerve binding peptides on sections ofhuman nerve showing high binding of HNP401. Exposure gain was decrease(30 to 10) for HNP401 as exposure under identical settings, compared toother standards, were saturated by high signal.

FIG. 4: Fluorescence from sectioned human nerve after topicalapplication of NP41, HNP401 and HNP404. Peptides were applied at 100 uMfor 20 mins followed by washing in PBS and imaging on a Nikon A1confocal microscope. All images are leveled equally.

FIG. 5: In-vivo labeling of Rat Sciatic nerve with HNP401

FIG. 6: Fluorescent labeling of rat prostate cavernosal nerve in liverats. HNP301 is an early generation nerve binding peptides that is notshowing as much contrast for prostate nerve labeling compared to HNP401.

FIG. 7: In-vivo labeling of prostatic neurovascular bundle with HNP401.HNP401 labeling of autonomic nerve bundles in live rats.

FIGS. 8A-8F: Screening of HUMAN nerve binding peptides identified byphage display. Topical application of 100 mM of human nerve bindingpeptides FAM-HNP401 (A), FAM-HNP402 (B), FAM-HNP403 (C) on serialsections of fresh-viable human sural nerve (top image) and humantemporalis muscle (bottom image). For comparison topical application of100 mM of carboxy-FAM (D) and peptide screened for binding to mousenerve NP41-FAM (E). H&E of staining of nerve and muscle (F). Allfluorescence images acquired on Lumar microscope at 34× magnificationwith a 2s exposure and levelled equally for comparison. NTQTLAKAPEHT(NP41; SEQ ID NO:15 from U.S. Pat. No. 8,685,372 or International PatentPublication No. WO2010121023A2).

FIGS. 9A-9H: Comparison of FAM-HNP401 and FAM-NP41 in binding andlabelling of HUMAN sural nerve. Topical application of 100 mM ofHNP401-FAM on 10 mm sections of unfixed human brachial plexus nervetissue (A) and human temporalis muscle tissue (D) kept adjacent on sameglass slide and imaged on a confocal microscope with 488 nm excitationlaser. For comparison, NP41-FAM was applied to human nerve (B) andmuscle (E) under identical conditions as mentioned for (A and D). H&Estaining of the nerve (C) and muscle (F). Signal intensity ofperineurium of nerve tissue treated with HNP401-FAM (n=4) compared withNP41-FAM (n=4) (G). Nerve to muscle contrast of peptides appliedtopically to human tissue sections (n=4) (H).

FIGS. 10A-10N: Differential binding of nerve binding peptides to HUMANand MOUSE tissue. Human tissue: Determination of optimal dose responseby topical application of HNP401-FAM on human sural nerve sections atfinal concentration of 375 mM (A), 100 mM (B), 50 mM (C), 10 mM (D) and1 mM (E), imaged with confocal microscopy with identical parameters andlevelled equally for comparison. ** brightened 2 fold for viewing. Nerveand muscle contrast at high concentration of 375 mM for FAM-NP41 (F andG) and FAM-HNP401 (H and I) imaged on confocal microscopy and levelledfor direct comparison. Mouse tissue: Mouse facial nerve (red arrows)with surrounding muscle treated with 375 mM (J), 100 mM (K) of FAM-NP41or 375 mM (1), 100 mM (M) of FAM-HNP401. Images in bottom row acquiredon Lumar imaging scope with identical parameters and are comparable.FAM-HNP401 shows high binding of muscle in mouse tissue with poorcontrast compared to FAM-NP41. Confocal imaging also showed thatHNP401-FAM binds epineurium, perineurium and endonerium but not axons(N).

FIGS. 11A-11H: In-vivo imaging of nerve binding peptides in RODENTS withpharmacokinetic profile following IV injection. In-vivo fluorescenceimage of sciatic nerve of 6 month old SKH1-Elite mice injected i.v. with450 nmols of FAM-HNP401 (A) or FAM-NP41 (B) and imaged on the Lumarimaging scope 2 h post injection. Intensity of sciatic nerve wasmeasured in Image J shows a 2.3 fold increase in binding for peptidescreened for binding human nerve (HNP401) vs peptide screened forbinding to mouse nerve (NP41) (C). However, the nerve to surroundingmuscle contrast for the two peptides are comparable at 5.79±0.81 forFAM-HNP401 and 6.63±1.63 for FAM-NP41 in mouse thigh (D). In-vivofluorescence image of prostate nerve plexus using real time customsurgical imaging system (E) and Lumar small animal microscope (F) 5hours after t.v. injection of 2 μmoles of HNP401-FAM in 100 gm maleSprague Dawley rat. Sciatic nerve in rat was imaged 5 hours aftersystemic injection of 2 μmoles of FAM-HNP401 (G). Blood clearance curveshows FAM signal obtained from equal volume of blood draws taken fromfive SKH1-Elite male mice. Each mouse was injected i.v. with 100 nmol ofFAM-HNP401 prior to blood collection at 1 min, 10 min, 20 min, 30 min, 1h and 2 h timepoints (H).

FIGS. 12A-12H: HNP401 binds to HUMAN nerves (cavernosal and medianante-brachial cutaneous) Fluorescent imaging after topical applicationof 100 μM FAM-HNP401 or FAM-NP41 on 10 μm sections on cryosectioningtape of nerve within human prostate gland, (top row, A and B) or frommedian anti-brachial cutaneous human nerve (bottom row, E and F). Nerveswere imaged immediately after sectioning and application of peptideusing confocal microscopy. Immunohistochemistry analysis with dual labelfor neurofilament antibody SMI312 (red) and DAPI stained nuclei (blue)(C and G) of fixed section of nerve and corresponding H&E staining (Dand H) on glass slides.

FIGS. 13A-13D: HNP401 binds to HUMAN cavernosal nerves. Topicalapplication of 100 μM FAM-HNP401 (A) or FAM-NP41 (B) on 10 μm sectionson cryosectioning tape of unfixed fresh viable nerves from the prostategland, using confocal microscopy. Immunofluorescence (C) neurofilamentantibody SMI312 on fixed section of nerve from prostate gland andcorresponding H&E staining (D) on glass slides. These images areobtained from different patients than those shown in FIG. 5. of thisdocument.

FIGS. 14A-141: Determination of HUMAN nerve binding domain of HNP401 bysequential deletion. Representative images fluorescence images ofunfixed human sural nerve treated topically with 100 uM of FAM labelledHNP401 (A), HNP401-N-2 (B), HNP401-N4 (C), HNP401-N6 (D), HNP401-N8 (E),HNP401-N4 C-2 (F), HNP401-N4 C-4 (G), HNP401-N4 C-6 (H), HNP401-N4 C-8(I).

FIG. 15: Quantitation of nerve binding of deletion variants of HNP401.Quantitation of nerve binding of each HNP401 variant shown in FIG. 7(n=5).

FIGS. 16A-16H: In-vivo fluorescent labelling of autonomic nerve inrodent. Low magnification fluorescent image showing bladder, vasdeferens and urethra running through the prostate with adjacentautonomic nerve labeled with FAM-NP41 in mice (A). Higher magnificationwhite light reflectance image (B) and corresponding fluorescencegrayscale image (C) of the autonomic nerve running adjacent to theurethra. Quantitation of autonomic nerve detection by fluorescencecompared to white light detection in mice (D) Nerve to muscle contrastfor reflectance/fluorescence were plotted for individual nerve branches.Values to the right of the line indicate that there is improvedvisualization with fluorescence compared to reflected light. Images(E-G) are analogous to (A-C) except they highlight FAM-NP41 dependentlabeling of autonomic nerve in rat prostate versus mouse, with whitelight imaging showing non-visible nerve (F). FAM-NP41 labeled prostatenerve is also detectable using a clinical grade Zeiss Pentero SurgicalMicroscope (H).

FIGS. 17A-17H: Comparison of FAM-HNP401 and FAM-NP41 in binding andlabelling of human sural nerve. Topical application of 100 μM ofFAM-HNP401 on 10 μm sections of unfixed human sural nerve tissue (A) andhuman temporalis muscle tissue (E) kept adjacent on same glass slide andimaged on a confocal microscope with 488 nm excitation laser. Forcomparison, FAM-NP41 was applied to a sequential section of human nerve(B) and muscle (F) under identical conditions as mentioned for (A andE). H&E staining of the nerve (C) and muscle (G). Signal intensity ofperineurium of nerve tissue treated with FAM-HNP401 (n=4) compared withFAM-NP41 (n=4) (D). Nerve to muscle contrast of peptides appliedtopically to human tissue sections (n=4) (H).

FIGS. 18A-18H: In-vivo imaging of nerve binding peptides in mouse andrat with pharmacokinetics. In-vivo fluorescence image of sciatic nervesfrom 6 month old SKH1 mice that had been previously intravenouslyinjected with 450 nmols of FAM-HNP401 [^(˜)48.4 mg/kg] (A) or FAM-NP41[^(˜)39 mg/kg] (B). Intensity of sciatic nerve measured and quantitatedin Image J showed a 2.3 fold increase for FAM-HNP401 compared toFAM-NP41 (C). Nerve to muscle contrast for the two peptides werecomparable at 5.79±0.81 for FAM-HNP401 and 6.63±1.63 for FAM-NP41 (D).In-vivo fluorescence image of rat sciatic nerve 5 hours afterintravenous injection of 2 μmoles of FAM-HNP401 [^(˜)54 mg/kg] (E). Ratprostate nerve imaged with real time custom surgical imaging system (F)and Lumar small animal microscope (G) 5 hours after intravenousinjection of 2 μmoles of FAM-HNP401. Blood clearance curve shows FAMsignal obtained from equal volume of blood draws taken from fiveSKH1-Elite male mice (H). Each mouse was injected intravenous with 100nmol [^(˜)11 mg/kg] of FAM-HNP401 prior to blood collection at 1 min, 10min, 20 min, 30 min, 1 h and 2 h time points.

FIGS. 19A-19H: HNP401 binds to fresh viable nerve from prostate glandand median anti-brachial cutaneous human nerve. Fluorescent imagingafter topical application of 100 μM FAM-HNP401 or FAM-NP41 on 10 μmsections on cryosectioning tape of nerve within human prostate gland,(top row, A and B) or from median anti-brachial cutaneous human nerve(bottom row, E and F). Nerves were imaged immediately after sectioningand application of peptide using confocal microscopy.Immunohistochemistry analysis with dual label for neurofilament antibodySMI312 (red) and DAPI stained nuclei (blue) (C and G) of fixed sectionof nerve and corresponding H&E staining (D and H) on glass slides.

FIGS. 20A-20K: Comparison of truncated sequences to determine bindingefficiency. Representative images fluorescence images of unfixed humansural nerve that were treated topically with 100 μM of FAM labelled N-2(A), N-4 (B), N-6 (C), N-8 (D), C-2 (E), C-4 (F), C-6 (G), C-8 (H) orHNP401 (I). Due to poor solubility C-6 had a final concentration of *73μM and C-8 had a final concentration of **80.6 μM for topical tests.Comparison of signal intensity of peptides normalized to FAM-HNP401 weremade to test for improved binding (J). Normalized sural nerve totemporalis muscle contrast was determined for FAM-HNP401 andFAM-HNP401-N-2 (Student t-test, unpaired, one-tail, p=0.011) (K).

FIGS. 21A-21C: Food dyes efficiently quench FAM-NP41 bladderfluorescence. A fluorescent plate reader assay was used to show dosedependent quenching of FAM-NP41 fluorescence. Erythrosine extra bluish(Santa Cruz Biotechnology, Inc.) was the most efficient quencher with>^(˜)80% quenching at 2.5 times dye to fluorescein ratio and >95%quenching at 5× dye to fluorescein ratio (A). Other food dyes testedincluded Allura Red and Sunset Yellow. To test for quenching in-vivo weadministered, by direct iv injection, 50 mg/kg MW 879.76 (^(˜)1.5 μmolesper 25 gm mouse) to mice that had been injected with 150 nmoles ofFAM-NP41 2 hours prior. This represents approximately a 10× dye toFAM-NP41 dose. Some bladder fluorescence remained after imaging soadditional dye (30 μl, 10 mM Erythrosin extra bluish) was injecteddirectly into the bladder. Images are shown for mouse bladder with nodye quencher (B) and addition of Erythrosine extra bluish (intravenousand intra bladder) (C) with bladder fluorescence quench to nearbackground level. Dye would likely not be needed if this method was usedfor human patients as bladder catheterization in patients could bestarted as FAM-NP41 is administered so bladder fluorescence could bewashed out.

FIGS. 22A-22E: TAMRA-NP41 labels autonomic unmyelinated nerve inprostate gland of rat. Fluorescence image of nerve fascicles (whitearrows) around prostate gland in living male Sprague-Dawley rat, imaged15 min post i.v. injection of 500 nmol of NP41-TAMRA (A). Tissue wasexcised and frozen unfixed for validation of peptide fluorescence signalusing confocal imaging (B) and immunohistochemistry with an antibody toTAMRA detected with horse radish peroxidase-secondary anddiaminobenzidine staining (C). Antibody staining against tyrosinehydroxylase was used to validate presence of autonomic nerves (D)no-primary negative control (E).

FIGS. 23A-23F: Screening of human nerve binding peptides identified byphage display. Topical application of 100 μM of human nerve bindingpeptides FAM-HNP401 (A), FAM-HNP402 (B), FAM-HNP403 (C) on serialsections of fresh-viable human sural nerve (upper row) and humantemporalis muscle (lower row). For comparison topical application of100M of carboxy-FAM (D) and peptide screened for binding to mouse nerveNP41-FAM (E). H&E of staining of nerve and muscle (F). All fluorescenceimages acquired on Lumar microscope at 34× magnification with a 2sexposure and levelled equally for comparison.

FIGS. 24A-24D: Screening of human nerve binding peptides identified byphage display. Topical application of 100 μM of human nerve bindingpeptides FAM-HNP401 (A), FAM-HNP402 (B), FAM-HNP403 (C) on serialsections of fresh-viable human ansa cervicalis nerve (upper row) andhuman great auricular nerve (lower row) from the neck of two differentpatients. For comparison topical application of 100 μM of carboxy-FAM(D) and peptide screened for binding to mouse nerve NP41-FAM (E). Allfluorescence images acquired on Lumar microscope at 34× magnificationwith a 2s exposure and levelled equally for comparison.

FIGS. 25A-25N: Differential binding of nerve binding peptides to humanand mouse tissue. Determination of optimal dose response by topicalapplication of FAM-HNP401 on human laryngeal nerve sections at finalconcentration of 375 μM (A), 100 μM (B), 50 μM (C), 10 μM (D) and 1 μM(E), imaged with confocal microscopy with identical parameters andlevelled equally for comparison. ** brightened 2 fold for viewing. Nerveto muscle contrast at high concentration of 375 μM for FAM-NP41 (F andG) and FAM-HNP401 (H and I) imaged on confocal microscopy and levelledfor direct comparison. Mouse facial nerve (red arrows) with surroundingmuscle treated with 375 μM (J), 100 μM (K) of FAM-NP41 or 375 μM (L),100 μM (M) of FAMHNP401. Images in bottom row acquired on Lumar imagingscope with identical parameters and are comparable. FAM-HNP401 showshigh binding of muscle in mouse tissue with poor contrast compared toFAMNP41. High resolution confocal image of low concentration ofFAM-HNP401 (10 μM) on human nerve shows binding of peptide to non-axonalstructural components of nerve (N).

FIGS. 26A-26B: Autofluorescence of human nerve tissue. Topicalapplication of 100 μM FAM-HNP401 (A) or buffer only (B) on 10 μmsections of unfixed human sural nerve followed by imaging using confocalmicroscopy under identical acquisition parameters for direct comparison.Images were levelled equally using Image J followed by a 16 foldbrightening of (B) for viewing.

FIGS. 27A-27C: Mass spectroscopy analysis of urine samples from miceinjected with nerve binding peptides. Fragmented ion peaks fromCysteine-FAM collected from the urine of mice that were injected withFAM-HNP401 indicating peptide is metabolized (A). Similar results wereobtained with mice injected with FAM-NP41 (B). However, mouse injectedwith FAM-dNP41, where peptide is made with d-amino acids, is detectablein the urine and is not metabolized (C).

FIGS. 28A-28F: Stability of peptides in ex-vivo human plasma andcerebrospinal fluid from rats. FAM-HNP401 peptide detected at 5 min (A)and 2 hours (B) after incubation at 37° C. in human plasma in at a doseof 53.2 mg/kg or 2 μmole. An equal volume of 1:1 acetonitrile:water with2% acetic acid is added to precipitate the protein matter, supernatantis extracted for analysis by LC-MS on a C18 reverse phase column withgradient of 9:1 H2O+0.05% TFA: Acetonitrile+0.05% TFA to 1:9 H2O+0.05%TFA:Acetonitrile+0.05% TFA in 20 min. Detector channel of 450 nm showsFAM-HNP401. The peptide remain intact at 2 hours post incubation with x% of the composition at 5 min post incubation with human plasma.FAM-NP41 peptide detected at 5 min (C) and 2 hours (D) after incubationat 37° C. in human plasma in at a dose of 53.2 mg/kg or 2 μmole,followed by LC-MS analysis with method described above. Similar to ourprevious result, FAM-NP41 remains intact at 2 hours post incubation withx % of the composition at 5 min post incubation with human plasma.FAM-HNP401 (E) and FAM-NP41 (F) were also tested in cerebrospinal spinalfluid from rat at 2 hours after incubation to demonstrate stability ofthe peptides in circulation.

FIGS. 29A-29D: FAM-HNP401 binds to fresh viable nerve acquired fromhuman prostate gland. Topical application of 100 μM FAM-HNP401 (A) orFAM-NP41 (B) on 10 μm sections of unfixed nerves from the prostate glandfollowed by imaging using confocal microscopy. Immunofluorescence fornerve using neurofilament antibody SMI312 (C) on fixed section of nervefrom prostate gland and corresponding H&E staining (D). These images areobtained from different patients than those shown in FIG. 20.

FIG. 30: Table of peptide sequences and their abbreviations.

DETAILED DESCRIPTION OF THE INVENTION I. Introduction

Disclosed herein, in certain embodiments, are targeting moleculescomprising a peptide that specifically binds to a human neuron, humannerve, or component of either. In some embodiments, the peptide isselected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16),Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4),Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments, the peptide is not Ac-SHSNTQTLAKAPEHTGC (Ac-NP41;SEQ ID NO:17). In some embodiments, the peptide is not SHSNTQTLAKAPEHTGC(NP41; SEQ ID NO:18). In some embodiments, the peptide is notNTQTLAKAPEHT (NP41; SEQ ID NO:19).

II. Definitions

In the present description, any concentration range, percentage range,ratio range, or integer range is to be understood to include the valueof any integer within the recited range and, when appropriate, fractionsthereof (such as one tenth and one hundredth of an integer), unlessotherwise indicated. Also, any number range recited herein relating toany physical feature, such as polymer subunits, size or thickness, areto be understood to include any integer within the recited range, unlessotherwise indicated. As used herein, the term “about” means±20% of theindicated range, value, or structure, unless otherwise indicated.

It should be understood that the terms “a” and “an” as used herein referto “one or more” of the enumerated components. The use of thealternative (e.g., “or”) should be understood to mean either one, both,or any combination thereof of the alternatives.

In addition, it should be understood that the individual compounds, orgroups of compounds, derived from the various combinations of thestructures and substituents described herein, are disclosed by thepresent application to the same extent as if each compound or group ofcompounds was set forth individually. Thus, selection of particularstructures or particular substituents is within the scope of the presentdisclosure

As used herein, the following terms have the meanings ascribed to themunless specified otherwise.

The central nervous system (CNS) consists of the brain and the spinalcord, as well as the retina.

The peripheral nervous system (PNS) extends outside the CNS. The PNS isdivided into the somatic nervous system and the autonomic nervoussystem.

A neuron is an electrically excitable cell that processes and transmitsinformation by electrical and chemical signaling.

A typical neuron possesses a cell body (often called the soma),dendrites, and an axon.

A nerve is an enclosed, cable-like bundle of neural axons. Each nerve isa cordlike structure that contains many axons. Each axon is surroundedby a layer of tissue called the endoneurium. The axons are bundledtogether into groups called fascicles, and each fascicle is wrapped in alayer of tissue called the perineurium. The neuron or nerve is wrappedin a layer of tissue called the epineurium.

As used herein, the term “targeting molecule” refers to any agent (e.g.,peptide, protein, nucleic acid polymer, aptamer, or small molecule) thatspecifically binds to a target of interest. In some embodiments, thetargeting molecule comprises a peptide, also referred to herein as“targeting peptide.” The target of interest may be a tissue, a celltype, a cellular structure (e.g., an organelle), a protein, a peptide, apolysaccharide, or a nucleic acid polymer. In some embodiments, thetargeting molecule is any agent that specifically binds to one or moreneurons or nerves of a subject. In some embodiments, the targetingmolecule further comprises a cargo (e.g., drug, fluorescent label, orphotosensitizing agent).

As used herein, the term “aptamer” refers to an oligonucleotide (e.g.,DNA, RNA, or XNA) molecule that has been selected from random poolsbased on their ability to bind other molecules with high affinityspecificity based on non-Watson and Crick interactions with the targetmolecule (see, e.g., Cox and Ellington, Bioorg. Med. Chem. 9:2525-2531(2001); Lee et al, Nuc. Acids Res. 32:D95-D100 (2004)) or a shortpeptide (e.g., 5-20 amino acids) that is embedded as a loop within astable protein scaffold rather than as a free peptide. Aptamers can beselected which bind nucleic acid, proteins, small organic compounds,vitamins, inorganic compounds, cells, and even entire organisms. In someembodiments, the targeting peptide can comprise an aptamer or thetargeting molecule peptide sequence can be in the format of an peptideaptamer.

The terms “polypeptide,” “peptide,” and “protein” are usedinterchangeably herein to refer to a polymer of amino acid residues. Theterms apply to naturally occurring occurring amino acid polymers as wellas amino acid polymers in which one or more amino acid residues is anon-naturally occurring amino acid (e.g., an amino acid analog). Theterms encompass amino acid chains of any length, including full lengthproteins (i.e., antigens), wherein the amino acid residues are linked bycovalent peptide bonds. As used herein, the term “peptide” refers to apolymer of amino acid residues typically ranging in length from 2 toabout 50 residues. In certain embodiments the peptide ranges in lengthfrom about 2, 3, 4, 5, 7, 9, 10, or 11 residues to about 50, 45, 40, 45,30, 25, 20, or 15 residues. In certain embodiments the peptide ranges inlength from about 8, 9, 10, 11, or 12 residues to about 15, 20 or 25residues. Where an amino acid sequence is provided herein, L-, D-, orbeta amino acid versions of the sequence are also contemplated as wellas retro, inversion, and retro-inversion isoforms. Peptides also includeamino acid polymers in which one or more amino acid residues is anartificial chemical analogue of a corresponding naturally occurringamino acid, as well as to naturally occurring amino acid polymers. Inaddition, the term applies to amino acids joined by a peptide linkage orby other modified linkages (e.g., where the peptide bond is replaced byan α-ester, a/3-ester, a thioamide, phosphonamide, carbamate,hydroxylate, and the like (see, e.g., Spatola, (1983) Chem. Biochem.Amino Acids and Proteins 7: 267-357), where the amide is replaced with asaturated amine (see, e.g., Skiles et al., U.S. Pat. No. 4,496,542,which is incorporated herein by reference, and Kaltenbronn et al.,(1990) Pp. 969-970 in Proc. 1 lth American Peptide Symposium, ESCOMScience Publishers, The Netherlands, and the like)).

The term “amino acid” refers to naturally occurring and synthetic aminoacids, as well as amino acid analogs and amino acid mimetics thatfunction in a manner similar to the naturally occurring amino acids.Naturally occurring amino acids are those encoded by the genetic code,as well as those amino acids that are later modified, e.g.,hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. An amino acidmay be an L- or D-amino acid. Amino acid analogs refer to compounds thathave the same basic chemical structure as a naturally occurring aminoacid, i.e., a carbon that is bound to a hydrogen, a carboxyl group, anamino group, and an R group, e.g., homoserine, norleucine, methioninesulfoxide. Such analogs have modified R groups (e.g., norleucine) ormodified peptide backbones, but retain the same basic chemical structureas a naturally occurring amino acid. Amino acid mimetics refers tochemical compounds that have a structure that is different from thegeneral chemical structure of an amino acid, but that functions in amanner similar to a naturally occurring amino acid.

Amino acids may be referred to herein by either their commonly knownthree letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, maybe referred to by their commonly accepted single-letter codes.

One of skill will recognize that individual substitutions, deletions oradditions to a peptide, polypeptide, or protein sequence which alters,adds or deletes a single amino acid or a small percentage of amino acidsin the encoded sequence is a “conservatively modified variant” where thealteration results in the substitution of an amino acid with achemically similar amino acid. Conservative substitution tablesproviding functionally similar amino acids are well known in the art.Such conservatively modified variants are in addition to and do notexclude polymorphic variants, interspecies homologs, and alleles of theinvention.

The following eight groups each contain amino acids that areconservative substitutions for one another: 1) Alanine (A), Glycine (G);2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine(Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L),Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y),Tryptophan (W); 7) Serine (S), Threonine (T); and 8) Cysteine (C),Methionine (M) (see, e.g., Creighton, Proteins (1984)).

“Sequence identity,” as used herein, refers to the percentage of aminoacid residues in a single sequence that are identical with the aminoacid residues in another reference polypeptide sequence after aligningthe sequences and introducing gaps, if necessary, to achieve the maximumpercent sequence identity, and not considering any conservativesubstitutions as part of the sequence identity. The percentage sequenceidentity values can be generated using the NCBI BLAST 2.0 software asdefined by Altschul et al. (1997), Nucl. Acids Res. 25:3389-3402, withthe parameters set to default values.

As used herein, the terms “label” refers to a molecule that facilitatesthe visualization and/or detection of a targeting molecule disclosedherein. In some embodiments, the label is a fluorescent moiety.

The phrase “specifically binds” when referring to the interactionbetween a targeting molecule disclosed herein and a target (e.g.,purified protein, neuron or nerve tissue, neuron or nerves, cranialneuron or nerves, central neuron or nerves, myelinated or unmyelinatedneuron or nerves, or connective tissue surrounding neuron or nerves),refers to the formation of a high affinity bond between the targetingmolecule and the target. Further, the term means that the targetingmolecule has low affinity for non-targets.

“Selective binding,” “selectivity,” and the like refer to the preferenceof agent to interact with one molecule as compared to another.Preferably, interactions between a targeting molecule disclosed hereinand a target are both specific and selective. Note that in someembodiments an agent is designed to “specifically bind” and “selectivelybind” two distinct, yet similar targets without binding to otherundesirable targets.

The terms “individual,” “patient,” or “subject” are usedinterchangeably. As used herein, they mean any mammal (i.e. species ofany orders, families, and genus within the taxonomic classificationanimalia: chordata: vertebrata: mammalia). In some embodiments, themammal is a cow, horse, sheep, pig, cat, dog, goat, mouse, rat, rabbit,guinea pig, non-human primate, or human. None of the terms require orare limited to situation characterized by the supervision (e.g. constantor intermittent) of a health care worker (e.g. a doctor, a registerednurse, a nurse practitioner, a physician's assistant, an orderly, or ahospice worker).

The terms “administer,” “administering”, “administration,” and the like,as used herein, refer to the methods that may be used to enable deliveryof agents or compositions to the desired site of biological action.These methods include, but are not limited to parenteral injection(e.g., intravenous, subcutaneous, intraperitoneal, intramuscular,intravascular, intrathecal, intravitreal, infusion, or local).Administration techniques that are optionally employed with the agentsand methods described herein, include e.g., as discussed in Goodman andGilman, The Pharmacological Basis of Therapeutics, current ed.;Pergamon; and Remington's, Pharmaceutical Sciences (current edition),Mack Publishing Co., Easton, Pa. In some embodiments, administration isvia systemic intravenous injection into human patients.

The term “pharmaceutically acceptable” as used herein, refers to amaterial that does not abrogate the biological activity or properties ofthe agents described herein, and is relatively nontoxic (i.e., thetoxicity of the material significantly outweighs the benefit of thematerial). In some instances, a pharmaceutically acceptable material maybe administered to an individual without causing significant undesirablebiological effects or significantly interacting in a deleterious mannerwith any of the components of the composition in which it is contained.

The term “surgery” as used herein, refers to any methods for that may beused to manipulate, change, or cause an effect by a physicalintervention. These methods include, but are not limited to opensurgery, endoscopic surgery, laparoscopic surgery, minimally invasivesurgery, robotic surgery, any procedures that may affect any neuron ornerves such as placement of retractors during spinal surgery, cardiacneuron or nerve ablation, epidural injection, intrathecal injections,neuron or nerve blocks, implantation of devices such as neuron or nervestimulators and implantation of pumps. In some embodiments, the subjectof the surgery is a human subject or human patient

III. Targets

Disclosed herein, in certain embodiments, are human neuron and/or nervetargeting molecules that specifically bind to a human neuron or nervetarget.

In some embodiments, the target is a human neuron or nerve. The nerve isany human nerve (e.g., motor nerves, sensory nerves, sympathetic andparasympathetic nerves, periprostatic neurovascular bundle, sciaticnerves, cranial nerves including olfactory nerve, optic nerve,oculomotor nerve, trochlear nerve, trigeminal nerve, abducens nerve,facial nerve, vestibulocochlear nerve, glossopharyngeal nerve, vagusnerve, accessory nerve, hypoglossal nerve, spinal nerves, brachialplexus, or lumbrosacral plexus). The neuron is any neuron (e.g., sensoryneurons (afferent neurons), motor neurons (efferent neurons),interneurons, unipolar neurons, bipolar neurons, multipolar neurons,basket cells, Betz cells, medium spiny neurons, Purkinje cells,pyramidal cells, Renshaw cells, Granule cells, anterior horn cells). Insome embodiments, the human neuron or nerve is myelinated. In someembodiments, the neuron or nerve is unmyelinated. In some embodiments,the human neuron or nerve is demyelinated. In some embodiments, thehuman neuron or nerve is undergoing demyelination.

In some embodiments, the neuron and/or nerve target is a component of ahuman neuron or nerve. The component of a human neuron or nerve is anycomponent of a neuron or nerve. In some embodiments, the target istissue within or surrounding a neuron or nerve (e.g., epineurium,perineurium, or endoneurium). In some embodiments, the target is acomponent of myelin, (e.g., myelin basic protein (MBP), myelinoligodendrocyte glycoprotein, or proteolipid protein). In someembodiments, the target is expressed by Schwann cells, (e.g., MBP, glialfibrillary acidic protein, S-100, or myelin protein zero). In someembodiments, the target is a component of neuron or nerve tissue, (e.g.,elastin, fibrillin, e-cadherin, cytokeratin, vimentin, collagen I,collagen, III, collagen IV, or collagen V). In some embodiments, thetarget is a neurotrophic factor receptor expressed in neuron or nerves,(e.g., tyrosine kinase receptors TrkA, TrkB, and TrkC, low affinityneuron or nerve growth receptor or p75 neurotrophin receptor, or GDNFfamily receptor alpha-1 or -2). In some embodiments, the target is anon-neurotrophic factor receptor expressed in a neuron or nerve tissue,(e.g., epithelial growth factor receptors, transforming growth factorbeta receptors, vascular endothelial growth factor receptors, endothelinA receptors, endothelin B receptors, and integrin receptors).

Determining whether a neuron and/or nerve targeting molecule is capableof binding a human neuron or nerve or component thereof is accomplishedby any suitable method. In some embodiments, the method of determiningwhether a neuron and/or nerve targeting molecule is capable of binding ahuman neuron or nerve or component thereof involves contacting atargeting molecule (e.g., peptide or aptamer) disclosed herein with atest agent for a period of time sufficient to allow the targetingmolecule and test agent to form a binding complex. The binding complexis detected using any suitable method. Suitable binding assays can beperformed in vitro or in vivo and include, but are not limited to, phagedisplay, two-hybrid screens, co-precipitation, cross-linking, andexpression cloning (see, e.g., Bennet, J. P. and Yamamura, H.I. (1985)“Neurotransmitter, Hormone or Drug Receptor Binding Methods,” inNeurotransmitter Receptor Binding (Yamamura, H. L, et al., eds.), pp.61-89. Other binding assays involve the use of mass spectrometry or NMRtechniques to identify molecules bound to the target of interest. Thetargeting molecule utilized in such assays can be naturally expressed,cloned or synthesized.

In some embodiments, the targeting molecule is capable of crossing theblood-brain barrier in order to reach and bind the human neuron or nerveof interest.

IV. Targeting Molecules Peptides and Aptamers

Provided in the present disclosure are peptides that bind to humanmotor/sensory and autonomic nerves and can be used in human neuron ornerve targeting molecules of the present invention. In some embodiments,a targeting peptide comprises an amino acid sequence ofSGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), or5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

In some embodiments, the human neuron or nerve targeting moleculecomprises a peptide sequence selected from the group consisting of:SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments, peptides of the present invention comprise an aminoacid sequence of about 8 to about 25 amino acids (e.g., 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids),about 10 to about 23 amino acids, or about 15 to about 21 amino acidscomprising the core binding domain of PYYVVKK (SEQ ID NO:40). In someembodiments, peptides of the present invention comprise an amino acidsequence of about 13 to about 25 amino acids comprising the core bindingdomain of PYYVVKK (SEQ ID NO:40) and an N-terminal sequence of QVPWEE(SEQ ID NO:41). In some embodiments, the peptides of the presentinvention comprise an amino acid core binding domain of PYY (SEQ IDNO:116) or PYYVV (SEQ ID NO:117) and an N-terminal sequence of QVPWEE(SEQ ID NO:41). In some embodiments, the peptides of the presentinvention comprise an amino acid core binding domain of PYY (SEQ IDNO:116) and an N-terminal sequence of QVPWEE (SEQ ID NO:41). In someembodiments, the peptides of the present invention comprise an aminoacid core binding domain of PYYVV (SEQ ID NO:117) and an N-terminalsequence of QVPWEE (SEQ ID NO:41).

One such embodiment is a peptide of QVPWEEPYYVVKK (SEQ ID NO:42). Insome embodiments, the targeting molecule comprises a peptide that is notAc-SHSNTQTLAKAPEHTGC (Ac-NP41 with GC linker; SEQ ID NO:17). In someembodiments, the targeting molecule comprises a peptide that is notSHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID NO:18). In someembodiments, the peptide is not NTQTLAKAPEHT (NP41; SEQ ID NO:19).

In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3).

In some embodiments, the targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS (HNP401-N-2;SEQ ID NO:20), and QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ IDNO:21).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1).

In some embodiments, the targeting molecule comprises the peptideWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2).

In some embodiments, the targeting molecule comprises the peptideDLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

In some embodiments, the targeting molecule comprises the peptideAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

In some embodiments, the targeting molecule comprises the peptideAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

In some embodiments, the targeting molecule comprises the peptideAc-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

In some embodiments, the targeting molecule comprises the peptideAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

In some embodiments, the targeting molecule comprises the peptideAc-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9).

In some embodiments, the targeting molecule comprises the peptideAc-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).

In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14).

In some embodiments, the targeting molecule comprises the peptideDTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16).

In some embodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).

In some embodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

In some embodiments, the targeting molecule comprises the peptidePWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22).

In some embodiments, the targeting molecule comprises the peptideEEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23).

In some embodiments, the targeting molecule comprises the peptidePYYVVKKSS (HNP401-N-8; SEQ ID NO:24).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEP (HNP401-C-8; SEQ ID NO:28).

In some embodiments, the targeting molecule comprises the peptidePWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118).

In some embodiments, the targeting molecule comprises the peptideEEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119).

In some embodiments, the targeting molecule comprises the peptidePYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123).

In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

In some embodiments, the targeting molecule comprises the peptide5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

In some embodiments, the targeting molecule comprises a peptide sequencesharing at least 80% homology with a peptide sequence disclosed herein.In some embodiments, the targeting molecule comprises a peptide sequencesharing at least 85% homology with a peptide sequence disclosed herein.In some embodiments, the targeting molecule comprises a peptide sequencesharing at least 90% homology with a peptide sequence disclosed herein.In some embodiments, the targeting molecule comprises a peptide sequencesharing at least 95% homology with a peptide sequence disclosed herein.In some embodiments, the targeting molecule comprises a peptide sequencesharing at least 99% homology with a peptide sequence disclosed herein.

In some embodiments, the targeting molecule comprises a peptide sequencehaving at least 75%, 80%, 85%, 90%, 95%, 97%, or 99% identity with apeptide sequence of SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSSGG (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), or5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

In some embodiments, the targeting molecule comprises an aptamer.

The peptides and aptamers of the present invention are synthesized byany suitable method. For example, targeting peptides and aptamers of thepresent invention can be chemically synthesized by solid phase peptidesynthesis. In some embodiments, peptides of the present invention areacetylated at the N-terminus (“Ac” or “acetyl”), amidated at theC-terminus (“CONH₂” or “NH₂”), or both. For example, the targetingpeptide may comprise Ac-SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:43),Ac-WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:44), Ac-DLPDIIWDFNWETA (HNP 403;SEQ ID NO:45), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ IDNO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:46), Ac-QVPWEEPYYVVKKSS(HNP401-N-2; SEQ ID NO:47), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGClinker; SEQ ID NO:7), Ac-QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker;SEQ ID NO:48), Ac-PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:49),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:50), Ac-EEPYYVVKKSSGGC (HNP401-N-6with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSS (HNP401-N-8; SEQ ID NO:51),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:52), Ac-SGQVPWEEPYYVVKKGGC(HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVV(HNP401-C-4; SEQ ID NO:53), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGClinker; SEQ ID NO:12), Ac-SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:54),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEP (HNP401-C-8; SEQ ID NO:55), Ac-SGQVPWEEPGGC (HNP401-C-8with GGC linker; SEQ ID NO:14), SGQVPWEEPYYVVKKSS-CONH₂ (HNP 401; SEQ IDNO:56), WEYHYVDLNWTSQHPQ-CONH₂ (HNP 402; SEQ ID NO:57),DLPDIIWDFNWETA-CONH₂ (HNP 403; SEQ ID NO:58), SGQVPWEEPYYVVKKSSGGC-CONH₂(HNP401 with GGC linker; SEQ ID NO:59), WEYHYVDLNWTSQHPQGGC-CONH₂(HNP402 with GGC linker; SEQ ID NO:60), DLPDIIWDFNWETAGGC-CONH₂ (HNP403with GGC linker; SEQ ID NO:61), DTHAHAKPRVPAFKSV-CONH₂ (HNP 404; SEQ IDNO:62), QVPWEEPYYVVKKSSGGC-CONH₂ (HNP401-N-2 with GGC linker; SEQ IDNO:63), QVPWEEPYYVVKKSSGG-CONH₂ (HNP401-N-2 with GG linker; SEQ IDNO:64), PWEEPYYVVKKSSGGC-CONH₂ (HNP401-N-4 with GGC linker; SEQ IDNO:65), EEPYYVVKKSSGGC-CONH₂ (HNP401-N-6 with GGC linker; SEQ ID NO:66),PYYVVKKSSGGC-CONH₂ (HNP401-N-8 with GGC linker; SEQ ID NO:67),SGQVPWEEPYYVVKKGGC-CONH₂ (HNP401-C-2 with GGC linker; SEQ ID NO:68),SGQVPWEEPYYVVGGC-CONH₂ (HNP401-C-4 with GGC linker; SEQ ID NO:69),SGQVPWEEPYYGGC-CONH₂ (HNP401-C-6 with GGC linker; SEQ ID NO:70),SGQVPWEEPGGC-CONH₂ (HNP401-C-8 with GGC linker; SEQ ID NO:71),QVPWEEPYYVVKKSS-CONH₂ (HNP401-N-2; SEQ ID NO:72), PWEEPYYVVKKSS-CONH₂(HNP401-N-4; SEQ ID NO:73), EEPYYVVKKSS-CONH₂ (HNP401-N-6; SEQ IDNO:74), PYYVVKKSS-CONH₂ (HNP401-N-8; SEQ ID NO:75),SGQVPWEEPYYVVKK-CONH₂ (HNP401-C-2; SEQ ID NO:76), SGQVPWEEPYYVV-CONH₂(HNP401-C-4; SEQ ID NO:77), SGQVPWEEPYY-CONH₂ (HNP401-C-6; SEQ IDNO:78), and SGQVPWEEP-CONH₂ (HNP401-C-8; SEQ ID NO:79),Ac-SGQVPWEEPYYVVKKSS-CONH₂ (HNP401; SEQ ID NO:80),Ac-WEYHYVDLNWTSQHPQ-CONH₂ (HNP402; SEQ ID NO:81),Ac-DLPDIIWDFNWETA-CONH₂ (HNP403; SEQ ID NO:82),Ac-SGQVPWEEPYYVVKKSSGGC-CONH₂ (HNP401 with GGC linker; SEQ ID NO:83),Ac-WEYHYVDLNWTSQHPQGGC-CONH₂ (HNP402 with GGC linker; SEQ ID NO:84),Ac-DLPDIIWDFNWETAGGC-CONH₂ (HNP403 with GGC linker; SEQ ID NO:85),Ac-DTHAHAKPRVPAFKSV-CONH₂ (HNP 404; SEQ ID NO:86),Ac-QVPWEEPYYVVKKSSGGC-CONH₂ (HNP401-N-2 with GGC linker; SEQ ID NO:87),Ac-QVPWEEPYYVVKKSSGG-CONH₂ (HNP401-N-2 with GG linker; SEQ ID NO:88),Ac-PWEEPYYVVKKSSGGC-CONH₂ (HNP401-N-4 with GGC linker; SEQ ID NO:89),Ac-EEPYYVVKKSSGGC-CONH₂ (HNP401-N-6 with GGC linker; SEQ ID NO:90),Ac-PYYVVKKSSGGC-CONH₂ (HNP401-N-8 with GGC linker; SEQ ID NO:91),Ac-SGQVPWEEPYYVVKKGGC-CONH₂ (HNP401-C-2 with GGC linker; SEQ ID NO:92),Ac-SGQVPWEEPYYVVGGC-CONH₂ (HNP401-C-4 with GGC linker; SEQ ID NO:93),Ac-SGQVPWEEPYYGGC-CONH₂ (HNP401-C-6 with GGC linker; SEQ ID NO:94),Ac-SGQVPWEEPGGC-CONH₂ (HNP401-C-8 with GGC linker; SEQ ID NO:95),Ac-QVPWEEPYYVVKKSS-CONH₂ (HNP401-N-2; SEQ ID NO:96),Ac-PWEEPYYVVKKSS-CONH₂ (HNP401-N-4; SEQ ID NO:97), Ac-EEPYYVVKKSS-CONH₂(HNP401-N-6; SEQ ID NO:98), Ac-PYYVVKKSS-CONH₂ (HNP401-N-8; SEQ IDNO:99), Ac-SGQVPWEEPYYVVKK-CONH₂ (HNP401-C-2; SEQ ID NO:100),Ac-SGQVPWEEPYYVV-CONH₂ (HNP401-C-4; SEQ ID NO:101), Ac-SGQVPWEEPYY-CONH₂(HNP401-C-6; SEQ ID NO:102), or Ac-SGQVPWEEP-CONH₂ (HNP401-C-8; SEQ IDNO:103). Techniques for solid phase synthesis are described, forexample, by Barany and Merrifield (1963) Solid-Phase Peptide Synthesis;pp. 3-284 in The Peptides: Analysis, Synthesis, Biology. Vol. 2: SpecialMethods in Peptide Synthesis, Part A.; Merrifield et al. (1963) J. Am.Chem. Soc, 85: 2149-2156, and Stewart et al. (1984) Solid Phase PeptideSynthesis, 2nd ed. Pierce Chem. Co., Rockford, 111.

V. Cargo

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a cargo. In some embodiments, a targeting peptidecomprises an amino acid sequence of SGQVPWEEPYYVVKKSS (HNP 401; SEQ IDNO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16),Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4),Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), or5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a cargo. In some embodiments, the human neuron ornerve targeting molecule comprises a peptide sequence selected from thegroup consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22), EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS(HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting of SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3). In some embodiments, the targeting molecule comprises apeptide selected from the group consisting of SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker;SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ IDNO:7), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS(HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2 with GGlinker; SEQ ID NO:21), and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 withGG linker; SEQ ID NO:104). In some embodiments, the targeting moleculecomprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In someembodiments, the targeting molecule comprises the peptideWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some embodiments, thetargeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3). In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In someembodiments, the targeting molecule comprises the peptideAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5). In someembodiments, the targeting molecule comprises the peptideAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In someembodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, thetargeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2with GG linker; SEQ ID NO:21). In some embodiments, the targetingmolecule comprises the peptide Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 withGGC linker; SEQ ID NO:7). In some embodiments, the targeting moleculecomprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In someembodiments, the targeting molecule comprises the peptideAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In someembodiments, the targeting molecule comprises the peptide EEPYYVVKKSS(HNP401-N-6; SEQ ID NO:23). In some embodiments, the targeting moleculecomprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQID NO:9). In some embodiments, the targeting molecule comprises thepeptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In some embodiments, thetargeting molecule comprises the peptide Ac-PYYVVKKSSGGC (HNP401-N-8with GGC linker; SEQ ID NO:10). In some embodiments, the targetingmolecule comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ IDNO:25). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ IDNO:11). In some embodiments, the targeting molecule comprises thepeptide SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26). In some embodiments,the targeting molecule comprises the peptide Ac-SGQVPWEEPYYVVGGC(HNP401-C-4 with GGC linker; SEQ ID NO:12). In some embodiments, thetargeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ IDNO:27). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). Insome embodiments, the targeting molecule comprises the peptide SGQVPWEEP(HNP401-C-8; SEQ ID NO:28). In some embodiments, the targeting moleculecomprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GGlinker; SEQ ID NO:104). In some embodiments, the targeting moleculecomprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQID NO:14). In some embodiments, the targeting molecule comprises thepeptide DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16). In some embodiments,the targeting molecule comprises a peptide that is notAc-SHSNTQTLAKAPEHTGC (Ac-NP41 with GC linker; SEQ ID NO:17). In someembodiments, the targeting molecule comprises a peptide that is notSHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID NO:18). In someembodiments, the peptide is not NTQTLAKAPEHT (NP41; SEQ ID NO:19).

In some embodiments, the peptide or aptamer is directly bound to acargo. In some embodiments, the peptide or aptamer is indirectly (e.g.,via a linker) bound to a cargo. In some embodiments, the peptide oraptamer is bound to a cargo at its N-terminus, at its C-terminus, or atan internal position (e.g., to an internal amino acid) of the peptide oraptamer. In some embodiments, two, three, four or more peptides oraptamers are directly or indirectly bound to a cargo. In certainembodiments, a cargo is a drug, fluorescent moeity, photosensitizingagent, or a combination thereof. In some embodiments, the cargo is adrug. In some embodiments, the cargo is a fluorescent moiety or afluorescent dye. In some embodiments, the cargo comprises a fluorescentmoiety or a fluorescent dye. In some embodiments, the cargo is aphotosensitizing agent. In some embodiments, the peptide or aptamer isbound to two or more cargo moieties. The two or more cargo moieties maybe the same moiety or different moieties, or be from the same class ofcargo moieties (e.g., two drugs) or from different classes of cargomoieties (e.g., one drug and one fluorescent moiety).

Common classes of fluorescent dyes include, but are not limited to,xanthenes such as rhodamines, rhodols and fluoresceins, and theirderivatives; bimanes; coumarins and their derivatives such asumbelliferone and aminomethyl coumarins; aromatic amines such as dansyl;squarate dyes; benzofurans; fluorescent cyanines; carbazoles;dicyanomethylene pyranes, polymethine, oxabenzanthrane, xanthene,pyrylium, carbostyl, perylene, acridone, quinacridone, rubrene,anthracene, coronene, phenanthrecene, pyrene, butadiene, stilbene,lanthanide metal chelate complexes, rare-earth metal chelate complexes,and derivatives of such dyes. Fluorescent dyes are discussed, forexample, in U.S. Pat. Nos. 4,452,720; 5,227,487; and 5,543,295.

In some embodiments, the fluorescent moiety or dye selected from thegroup consisting of a xanthene; a bimane; a coumarin; an aromaticamines; a benzofuran; a fluorescent cyanine; a carbazole; adicyanomethylene pyrane; polymethine; oxabenzanthrane; pyrylium;carbostyl; perylene; acridone; quinacridone; rubrene; anthracene;coronene; phenanthrecene; pyrene; butadiene; stilbene; porphyrin;pthalocyanine; lanthanide metal chelate complexes; rare-earth metalchelate complexes; FITC; Cy3; EGFP; cyan fluorescent protein (CFP);EGFP; 5-FAM; 6-FAM; FAM; fluorescein, IAEDANS, EDANS and BODIPY FL;TRITC; Cy5; Cy3; YFP; 6-FAM; LC Red 640; Alexa Fluor 546; fluorescein;tetramethylrhodamine; Dabcyl; BODIPY FL; QSY 7, QSY 9, QSY 21 andBBQ-650 dyes.

In some embodiments, the cargo comprises fluorescein dyes. Typicalfluorescein dyes include, but are not limited to, 5-carboxyfluorescein,fluorescein-5-isothiocyanate, 5(6)-carboxyfluorescein,5,6-dicarboxyfluorescein, 5-(and 6)-sulfofluorescein,sulfonefluorescein, succinyl fluorescein, 5-(and 6)-carboxy SNARF-1,carboxyfluorescein sulfonate, carboxyfluorescein zwitterion,carboxyfluorescein quaternary ammonium, carboxyfluorescein phosphonate,carboxyfluorescein GABA, carboxyfluorescein-cys-Cy5,5′(6′)-carboxyfluorescein, fluorescein glutathione, and6-carboxyfluorescein; examples of other fluorescein dyes can be found,for example, in U.S. Pat. Nos. 6,008,379, 5,750,409, 5,066,580, and4,439,356. A cargo may include a rhodamine dye, such as, for example,5-(and 6)-carboxy rhodamine 110, tetramethylrhodamine-6-isothiocyanate,5-carboxytetramethylrhodamine, 5-carboxy rhodol derivatives, tetramethyland tetraethyl rhodamine, diphenyldimethyl and diphenyldiethylrhodamine, dinaphthyl rhodamine, rhodamine 101 sulfonyl chloride (soldunder the tradename of TEXAS RED®), and other rhodamine dyes. Otherrhodamine dyes can be found, for example, in U.S. Pat. Nos. 6,080,852;6,025,505; 5,936,087; 5,750,409. In some embodiments, a cargo moietyincludes a cyanine dye, such as, for example, Cy3, Cy3B, Cy3.5, Cy5,Cy5.5, Cy 7.

In some embodiments, cargo moiety comprises fluorophores. Fluorophoresare commercially available and any known and/or commercially availablefluorophore can be employed as the cargo. In some embodiments, thefluorophore exhibits green fluorescence (such as for example 494 nm/519nm), orange fluorescence (such as for example 554 nm/570 nm), redfluorescence (such as for example 590 nm/617 nm), or far redfluorescence (such as for example 651 nm/672 nm) excitation/emissionspectra. In some embodiments, the fluorophore is a fluorophore withexcitation and emission spectra in the range of about 350 nm to about775 nm. In some embodiments the excitation and emission spectra areabout 346 nm/446 nm, about 494 nm/519 nm, about 554 nm/570 nm, about 555nm/572 nm, about 590 nm/617 nm, about 651 nm/672 nm, about 679 nm/702 nmor about 749 nm/775 nm. In some embodiments, the fluorophore can includebut is not limited to AlexaFluor 3, AlexaFluor 5, AlexaFluor 350,AlexaFluor 405, AlexaFluor 430, AlexaFluor 488, AlexaFluor 500,AlexaFluor 514, AlexaFluor 532, AlexaFluor 546, AlexaFluor 555,AlexaFluor 568, AlexaFluor 594, AlexaFluor 610, AlexaFluor 633,AlexaFluor 647, AlexaFluor 660, AlexaFluor 680, AlexaFluor 700, andAlexaFluor 750 (Molecular Probes AlexaFluor dyes, available from LifeTechnologies, Inc. (USA)). In some embodiments, the fluorophore caninclude but is not limited to Cy dyes, including Cyt, Cy3, Cy3B, Cy3.5,Cy5, Cy5.5 and Cy7 (available from GE Life Sciences or Lumiprobes). Insome embodiments the fluorophore can include but is not limited toDyLight 350, DyLight 405, DyLight 488, DyLight 550, DyLight 594, DyLight633, DyLight 650, DyLight 680, DyLight 750 and DyLight 800 (availablefrom Thermo Scientific (USA)). In some embodiments, the fluorophore caninclude but is not limited to a FluoProbes 390, FluoProbes 488,FluoProbes 532, FluoProbes 547H, FluoProbes 594, FluoProbes 647H,FluoProbes 682, FluoProbes 752 and FluoProbes 782, AMCA, DEAC(7-Diethylaminocoumarin-3-carboxylic acid);7-Hydroxy-4-methylcoumarin-3; 7-Hydroxycoumarin-3; MCA(7-Methoxycoumarin-4-acetic acid); 7-Methoxycoumarin-3; AMF(4′-(Aminomethyl)fluorescein); 5-DTAF(5-(4,6-Dichlorotriazinyl)aminofluorescein); 6-DTAF(6-(4,6-Dichlorotriazinyl)aminofluorescein); FAM; 6-FAM(6-Carboxyfluorescein), 5(6)-FAM cadaverine; 5-FAM cadaverine; 5(6)-FAMethylenediamme; 5-FAM ethylenediamme; 5-FITC (FITC Isomer I;fluorescein-5-isothiocyanate); 5-FITC cadaverin;Fluorescein-5-maleimide; 5-IAF (5-lodoacetamidofluorescein); 6-JOE(6-Carboxy-4′,5′-dichloro-2′,7′-dimethoxyfluorescein); 5-CRI 10(5-Carboxyrhodamine 110); 6-CRI 10 (6-Carboxyrhodamine 110); 5-CR6G(5-Carboxyrhodamine 6G); 6-CR6G (6-Carboxyrhodamine 6G);5(6)-Carboxyrhodamine 6G cadaverine; 5(6)-Caroxyrhodamine 6Gethylenediamme; 5-ROX (5-Carboxy-X-rhodamine); 6-ROX(6-Carboxy-X-rhodamine); 5-TAMRA (5-Carboxytetramethylrhodamine);6-TAMRA (6-Carboxytetramethylrhodamine); 5-TAMRA cadaverine; 6-TAMRAcadaverine; 5-TAMRA ethylenediamme; 6-TAMRA ethylenediamme; 5-TMR C6maleimide; 6-TMR C6 maleimide; TR C2 maleimide; TR cadaverine; 5-TRITC;G isomer (Tetramethylrhodamine-5-isothiocyanate); 6-TRITC; R isomer(Tetramethylrhodamine-6-isothiocyanate); Dansyl cadaverine(5-Dimethylaminonaphthalene-1-(N-(5-aminopentyl))sulfonamide); EDANS C2maleimide; fluorescamine; NBD; and pyrromethene and derivatives thereof.

In some embodiments, a cargo comprises an environmentally sensitivefluorescent dye or fluorophore. Examples of environmentally sensitivefluorescent dyes or fluorophores include 5,6-carboxy-diethyl rhodol (pHsensitive), merocyanine (membrane potential sensitive), and Nile redcarboxylic acid (lipid sensitive).

In some embodiments, a cargo comprises a photosensitizing agent. Aphotosensitizing agent is any agent or compound useful in light inducedablation therapy. Such agents, when exposed to a specific wavelength oflight, react with molecular oxygen to produce singlet oxygen, which ishighly cytotoxic. Thus, targeting molecules of the present inventioncomprising a photosensitizing agent may be used to focally injurenerves. In certain embodiments, a photosensitizing agent is a porphyrin,chlorin, or dye. Examples of photosensitizing agents include porphyrin,protoporfin IX, purlytin, verteporfin, HPPH, temoporfin, methylene blue,photofrin, protofrin, hematoporphyrin, Talaporfin, benzopophyrinderivative monoacid, 5-aminileuvolinic acid, Lutetium texaphyrin,metallophthalocyanine, metallo-naphthocyaninesulfobenzo-porphyrazines,metallo-naphthalocyanines, zinc tetrasulfophthalocyanine,bacteriochlorins, metallochlorins, chlorine derivative,Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein(DBF), IR700DX, naphthalocyanine, and porphyrin derivatives. In someembodiments, the photosensitizing agent is conjugated to a C-terminalcysteine residue of the human neuron or nerve targeting molecule viamaleimide mediated conjugation. Preferably, the photosensitizing agentof the present invention is activated by light having a wavelength ofbetween 400 nm to 700 nm. Still more preferably, the photosensitizingagent in the present invention is activated at 627 nm and 660 nm. Anoptimal light dose can be identified to generate maximal nerve killingwith minimal injury to adjacent tissue.

VI. Drugs

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a drug. All drugs that act on a neuron or nerve (or acomponent thereof) are encompassed within the term “drug.” Specificexamples of drug given herein, are illustrative and are not meant tolimit the drugs for use with the targeting molecules disclosed herein.In some embodiments, the peptide or aptamer is directly bound to a drug.In some embodiments, the peptide or aptamer is indirectly (e.g., via alinker) bound to a drug. In some embodiments, two or more peptides oraptamers are directly or indirectly bound to a drug. In someembodiments, the human neuron or nerve targeting molecule furthercomprises a cargo. In some embodiments, the human neuron or nervetargeting molecule comprises a peptide sequence selected from the groupconsisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), or5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3). In some embodiments, the targeting molecule comprises apeptide selected from the group consisting of SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker;SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ IDNO:7), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS(HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2 with GGlinker; SEQ ID NO:21), and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 withGG linker; SEQ ID NO:104). In some embodiments, the targeting moleculecomprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In someembodiments, the targeting molecule comprises the peptideWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some embodiments, thetargeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3). In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In someembodiments, the targeting molecule comprises the peptideAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5). In someembodiments, the targeting molecule comprises the peptideAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In someembodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, thetargeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2with GG linker; SEQ ID NO:21). In some embodiments, the targetingmolecule comprises the peptide Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 withGGC linker; SEQ ID NO:7). In some embodiments, the targeting moleculecomprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In someembodiments, the targeting molecule comprises the peptideAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In someembodiments, the targeting molecule comprises the peptide EEPYYVVKKSS(HNP401-N-6; SEQ ID NO:23). In some embodiments, the targeting moleculecomprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQID NO:9). In some embodiments, the targeting molecule comprises thepeptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In some embodiments, thetargeting molecule comprises the peptide Ac-PYYVVKKSSGGC (HNP401-N-8with GGC linker; SEQ ID NO:10). In some embodiments, the targetingmolecule comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ IDNO:25). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ IDNO:11). In some embodiments, the targeting molecule comprises thepeptide SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26). In some embodiments,the targeting molecule comprises the peptide Ac-SGQVPWEEPYYVVGGC(HNP401-C-4 with GGC linker; SEQ ID NO:12). In some embodiments, thetargeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ IDNO:27). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). Insome embodiments, the targeting molecule comprises the peptide SGQVPWEEP(HNP401-C-8; SEQ ID NO:28). In some embodiments, the targeting moleculecomprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQID NO:14). In some embodiments, the targeting molecule comprises thepeptide DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16). In some embodiments,the targeting molecule comprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104).

In some embodiments, the drug is selected from a drug that: induces celldeath (apoptotic or necrotic), inhibits cell death (apoptotic ornecrotic), inhibits the transmission of a neuron or nerve signal (i.e.,an electrochemical impulse), inhibits the release of a neurotransmitter,agonizes the activity of a GABA receptor, partially or fully inhibitsthe repolarization of a neuron, disrupts the conduction of an ionchannel, or a combination thereof.

In some embodiments, the drug is an antihistamine, a GABA receptormodulator, a neurotransmitter reuptake inhibitor, a local anesthetic, ananticholinergic, a sodium channel blocker, a calcium channel blocker, athyrotropin-releasing hormone, a γ-secretase inhibitor, an AMPA receptoragonist or antagonist, an NMDA receptor agonist or antagonist, an mGlureceptor agonist or antagonist, a growth factor, an antiemetic agent, acorticosteroid; a cytotoxic agent; an antioxidant, an iron chelator, amitochondrial modulator, a sirtuin modulator, a nitric oxide (NO) and/ornitric oxide synthase (NOS) modulator, a potassium channel agonist orantagonist, a purigenic receptor agonist or antagonist, or a combinationthereof.

In some embodiments, the drug is meclizine, diphenhydramine,dimenhydrinate, loratadine, quetiapine, mepyramine, piperoxan,antazoline, carbinoxamine, doxylamine, clemastine, pheniramine,chlorphenamine, chlorpheniramine, dexchlorpheniramine, brompheniramine,triprolidine, cyclizine, chlorcyclizine, hydroxyzine, promethazine,alimemazine, trimeprazine, cyproheptadine, azatadine, ketotifen,oxatomide, meclizine hydrochloride, promethazine hydrochloride,cinnarizine, hydroxyzine pamoate, betahistine dihydrochloride,alprazolam, bromazepam, brotizolam, chlordiazepoxide, clonazepam,clorazepate, diazepam, estazolam, flunitrazepam, flurazepam, loprazolam,lorazepam, lormetazepam, idazolam, nimetazepam, nitrazepam, oxazepam,prazepam, temazepam, triazolam, clonazepam, diazepam, lorazepam,furosemide, bumetanide, ethacrynic acid, gabapentin, pregabalin,muscimol, baclofen, amitriptyline, nortriptyline, trimipramine,fluoxetine, paroxetine, sertraline, glycopyrrolate, homatropine,scopolamine, atropine, benzocaine, carticaine, cinchocaine,cyclomethycaine, lidocaine, prilocaine, propxycaine, proparacaine,tetracaine, tocainide, trimecaine, carbamazepine, oxcarbazepine,phenytein, valproic acid, sodium valproate, cinnarizine, flunarizine,nimodipine, thyrotropin-releasing hormone, amifostine (also known asWR-2721, or ETHYOL®); a carbamate compound (e.g.,2-phenyl-1,2-ethanediol monocarbomates and dicarbamates); LY450139(hydroxylvaleryl monobenzocaprolactam); L685458(IS-benzyl-4R[I-[I-S-carbamoyl-2-phenethylcarbamoyl)-IS-3-methylbutylcarbamoyl]-2R-hydroxy-5-phenylpentyl}carbamicacid tert-butyl ester); LY411575(N2-[(2S)-2-(3,5-difluorophenyl)-2-hydroxyethanoyl]-N1[(7S)-5-methyl-6-oxo-6,7-dihydro-5H-dibenzo[bid]azepin-7yl]-L-alaninamide);MK-0752; tarenflurbil; BMS-299897 (2-[(IR)-1-[[(4-chlorophenyl)sulfony](2,5-difluorophenyl)amino]ethyl]-5-fluorobenzenepropanoic acid;CNQX (6-cyano-7-nitroquinoxaline-2,3-dione); NBQX(2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f]quinoxaline-2,3-dione); DNQX(6,7-dinitroquinoxaline-2,3-dione); kynurenic acid;2,3-dihydroxy-6-nitro-7-sulfamoylbenzo-[f]quinoxaline;1-aminoadamantane; dextromethorphan; dextrorphan; ibogaine; ketamine;nitrous oxide; phencyclidine; riluzole; tiletamine; memantine;dizocilpine; aptiganel; remacimide; 7-chlorokynurenate; DCKA(5,7-dichlorokynurenic acid); kynurenic acid;1-aminocyclopropanecarboxylic acid (ACPC); AP7(2-amino-7-phosphonoheptanoic acid); APV(R-2-amino-5-phosphonopentanoate); CPPene(3-[(R)-2-carboxypiperazin-4-yl]-prop-2-enyl-1-phosphonic acid);(+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-pro-panol;(1 S,2S)-1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenylpiperi-dino)-1-propanol;(3R,4S)-3-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl-)-chroman-4;7-diol;(IR*,2R*)-1-(4-hydroxy-3-memylphenyl)-2-(4-(4-fluoro-phenyl)-4-hydroxypiperidin-1-yl)-propan-1-ol-mesylate);LY389795 ((−)-2-thia-4-aminobicyclo-hexane-4,6-dicarboxylate); LY379268((−)-2-oxa-4-aminobicyclo-hexane-4,6-dicarboxylate); LY354740((+)-2-aminobicyclo-hexane-2,6dicarboxylate); DCG-IV((2S,2′R,3′R)-2-(2′,31-dicarboxycyclopropyl)glycine); 2R,4R-APDC(2R,4R-4-aminopyrrolidine-2,4-dicarboxylate); (S)-3C4H PG((S)-3-carboxy-4-hydroxyphenylglycine); (S)-4C3HPG((S)-4-carboxy-3-hydroxyphenylglycine); L-CCG-I((2S,1′S,2′S)-2-(carboxycyclopropyl)glycine); ACPT-I((1S,3R,4S)-1-aminocyclopentane-1,3,4-tricarboxylic acid); L-AP4(L-(+)-2-Amino-4-phosphonobutyric acid); (S)-3,4-DCPG((S)-3,4-dicarboxyphenylglycine); (RS)-3,4-DCPG ((RS)-3,4-dicarboxyphenylglycine); (RS)-4-phosphonophenylglycine ((RS)PPG); AMN082(,N′-bis(diphenylmethyl)-1,2-ethanediamine dihydrochloride); DCG-IV((2S,2′R,3′R)-2-(2′,3′-dicarboxycyclopropyl)glycine); AMN082;brain-derived neurotrophic factor (BDNF); ciliary neurotrophic factor(CNTF); glial cell-line derived neurotrophic factor (GDNF);neurotrophin-3; neurotrophin-4; fibroblast growth factor (FGF) receptor;insulin-like growth factor (IGF); an aminoglycoside antibiotic (e.g.,gentamicin and amikacin); a macrolide antibiotic (e.g, erythromycin); aglycopeptide antibiotic (e.g. vancomycin); salicylic acid; nicotine;Eburnamenine-14-carboxylic acid ethyl ester; sipatrigine(2-(4-Methylpiperazin-1-yl)-5-(2,3,5-trichlorophenyl)-pyrimidin-4-amine);amiloride (3,5-diamino-N-(aminoiminomethyl)-6-chloropyrazinecarbox amidehydrochloride); carbamazepine (5H-dibenzo[b,f]azepine-5-carboxamide);TTX(octahydro-12-(hydroxymethyl)-2-imino-5,9:7,10a-dimethano-10aH-[1,3]dioxocino[6,5-d]pyrimidine-4,7,10,11,12-pentol);RS100642 (1-(2,6-dimethyl-phenoxy)-2-ethylaminopropane hydrochloride);mexiletine ((1-(2,6-dimethylphenoxy)-2-aminopropane hydrochloride));QX-314 (N-(2,6-Dimethylphenylcarbamoylmethyl)triethylammonium bromide);phenytoin (5,5-diphenylimidazolidine-2,4-dione); lamotrigine(6-(2,3-dichlorophenyl)-1,2,4-triazine-3,5-diamine); 4030W92(2,4-diamino-5-(2,3-dichlorophenyl)-6-fluoromethylpyrimidine); BW1003C87(5-(2,3,5-trichlorophenyl) pyrimidine-2,4-1.1 ethanesulphonate); QX-222(2-[(2,6-dimethylphenyl)amino]-N,N,N-trimethyl-2-oxoetha niminiumchloride); ambroxol (trans-4-[[(2-Amino-3, 5-dibromophenyl)methyl]amino] cyclo hexanol hydrochloride); R56865(N-[1-(4-(4-fluorophenoxy)butyl]-4-piperidinyl-N-methyl-2-benzo-thiazolamine);lubeluzole; ajmaline ((17R,21alpha)-ajmalan-17,21-diol); procainamide(4-amno-N-(2-diethylaminoethyljbenzamide hydrochloride); flecainide;riluzoleor; triamicinolone actenoide; Dexamethasone; promethazine;prochlorperazine; trimethobenzamide; triethylperazine; dolasetron;granisetron; ondansetron; tropisetron; and palonosetron; droperidol;meclizine; perphenazine; thiethyl perazine; domperidone; properidol;haloperidol; chlorpromazine; promethazine; prochlorperazine;metoclopramide; dronabinol; nabilone; sativex; scopolamine;dexamethasone; trimethobenzamine; emetrol; propofol; muscimol; acridinecarboxamide; actinomycin; 17-N-allylamino-17-demethoxygeldanamycin;amsacrine; aminopterin; anthracycline; antineoplastic; antineoplaston;5-azacytidine; azathioprine; BL22; bendamustine; biricodar; bleomycin;bortezomib; bryostatin; busulfan; calyculin; camptothecin; capecitabine;carboplatin; chlorambucil; cisplatin; cladribine; clofarabine;cytarabine; dacarbazine; dasatinib; daunorubicin; decitabine;dichloroacetic acid; discodermolide; docetaxel; doxorubicin; epirubicin;epothilone; eribulin; estramustine; etoposide; exatecan; exisulind;ferruginol; floxuridine; fludarabine; fluorouracil; fosfestrol;fotemustine; gemcitabine; hydroxyurea; IT-101; idarubicin; ifosfamide;imiquimod; irinotecan; irofulven; ixabepilone; laniquidar; lapatinib;lenalidomide; lomustine; lurtotecan; mafosfamide; masoprocol;mechlorethamine; melphalan; mercaptopurine; mitomycin; mitotane;mitoxantrone; nelarabine; nilotinib; oblimersen; oxaliplatin; PAC-I;methotrexate (RHEUMATREX®, Amethopterin); cyclophosphamide (CYTOXAN®)thalidomide (THALID OMID®); paclitaxel; pemetrexed; pentostatin;pipobroman; pixantrone; plicamycin; procarbazine; proteasome inhibitors(e.g.; bortezomib); raltitrexed; rebeccamycin; rubitecan; SN-38;salinosporamide A; satraplatin; streptozotocin; swainsonine; tariquidar;taxane; tegafur-uracil; temozolomide; testolactone; thioTEPA;tioguanine; topotecan; trabectedin; tretinoin; triplatin tetranitrate;tris(2-chloroethyl)amme; troxacitabine; uracil mustard; valrubicin;vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar;N-acetylcysteine; vitamin E; vitamin C; vitamin A; lutein; seleniumglutathione; melatonin; a polyphenol; a carotenoid; coenzyme Q-10;Ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one (also called PZ 51 orDR3305); L-methionine; azulenyl nitrones; L-(+)-Ergothioneine; CAPE(caffeic acid phenethyl ester); dimethylthiourea; dimethylsulfoxide;disufenton sodium; pentoxifylline; MCI-186; Ambroxol; U-83836E; MitoQ(mitoquinone mesylate); Idebenone(2-(10-hydroxydecyl)-5,6-dimethoxy-3-methyl-cyclohexa-2,5-diene-1,4-dione);desferrioxamine; hydroxybenzyl ethylene diamine; fullerenol-1,pyrrolidine dithiocarbamate; acetylcarnitine; lipoic acid; a stilbene; achalcone; a flavone; an isoflavone; a flavanones; an anthocyanidin; acatechin; isonicotinamide; dipyridamole; ZM 336372; camptothecin;coumestrol; nordihydroguaiaretic acid; esculetin; SRT-1720; SRT-1460;SRT-2183; aminoguanidine;1-Amino-2-hydroxyguanidine p-toluensulfate;GED; bromocriptine mesylate; dexamethasone; SDMA; ADMA; L-NMMA; L-NMEA;D-MMA; L-NIL; L-NNA; L-NPA; L-NAME; L-VNIO; diphenyleneiodoniumchloride; 2-ethyl-2-thiopseudourea; haloperidol; L-NIO; MEG; SMT; SMTC;7-Ni; nNOS inhibitor; 1,3-PBITU; L-thiocitrulline; TRIM; MTR-105; BBS-I;BBS-2; ONO-1714; GW273629; GW 274150; PPA250; AR-R17477; AR-R18512;spiroquinazolone; 1400W; S-NC; NTG; SNP; thapsigargin; VEGF; bradykinin;ATP; sphingosine-1-phosphate; estrogen; angiopoietin; acetylcholine;SIN-I; GEA 3162; GEA; GEA 5024; GEA 5538; SNAP; molsidomine; CNO-4;CNO-5; DEA/NO; IPA/NO; SPER/NO; SULFI/NO; OXI/NO; DETA/NO; nicorandil;minoxidil, levcromakalim; lemakalim; cromakalim; L-735,334; retigabine;flupirtine; BMS-204352; DMP-543; linopirdine; XE991; 4-AP; 3,4-DAP;E-4031; DIDS; Way 123,398; CGS-12066 A; dofetilide; sotalol; apamin;amiodarone; azimilide; bretylium; clofilium; tedisamil; ibutilide;sematilide; nifekalant; tamulustoxin; ATP; ADP; UTP; UDP; UDP-glucose;adenosine; 2-MESATP; 2-MESADP; ABMEATP; DATPAS; ATPrS; BZ-ATP; MRS2703;DENUFOSOL TETRASODIUM; MRS2365; MRS 2690; PSB 0474; A-317491; RO-3(Roche); SURAMIN; PPADS; PPNDS; DIDS; pyridoxal-5-phosphate;5-(3-bromophenyl)-1,3-dihydro-2H-benzofuro-[3,2-e]-1,4-diazepin-2-one;cibacron blue; basilen blue; ivermectin; A-438079; A-740003; NF023;NF449; NFI 10; NF157; MRS 2179; NF279; MRS 2211; MRS 2279; MRS 2500tetrasodium salt; TNP-ATP; tetramethylpyrazine; Ip51;jQy-carboxymethylene ATP; βγ-chlorophosphomethylene ATP; KN-62;spinorphin; minocycline; SB-203580(4-(4-Fluorophenyl)-2-(4-methylsulfmyl phenyl)-5-(4-pyridyl)1H-imidazole); PD 169316(4-(4-Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-1H-imidazole); SB202190(4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole); RWJ67657(4-[4-(4-fluorophenyl)-1-(3-phenylpropyl)-5-(4-pyridinyl)-1H-imidazol-2-yl]-3-butyn-1-ol); SB 220025(5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinlyl)imidazole);D-JNKI-I ((D)-hJIPi75-i57-DPro-DPro-(D)-HIV-T AT57-48); AM-111 (Auris);SP600125 (anthra[1,9-cd]pyrazol-6(2H)-one); JNK Inhibitor I((L)-HIV-TAT48-57-PP-JBD20); JNK Inhibitor III((L)-HIV-TAT47-57-gaba-c-Jun633-57); AS601245 (1,3-benzothiazol-2-yl(2-[[2-(3-pyridinyl) ethyl] amino]-4 pyrimidinyl) acetonitrile); JNKInhibitor VI (H2N-RPKRPTTLNLF-NH2); JNK Inhibitor VIII(N-(4-Amino-5-cyano-6-ethoxypyridin-2-yl)-2-(2,5-dimethoxyphenyl)acetamide);JNK Inhibitor IX(N-(3-Cyano-4,5,6,7-tetrahydro-1-benzothien-2-yl)-1-naphthamide);dicumarol (3,3′-Methylenebis(4-hydroxycoumarin)); SC-236(4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yljbenzene-sulfonamide);CEP-1347 (Cephalon); CEP-11004 (Cephalon); an artificial proteincomprising at least a portion of a Bcl-2 polypeptide; a recombinant FNK;V5 (also known as Bax inhibitor peptide V5); Bax channel blocker((±)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol); Baxinhibiting peptide P5 (also known as Bax inhibitor peptide P5); Kp7-6;FAIM(S) (Fas apoptosis inhibitory molecule-short); FAIM(L) (Fasapoptosis inhibitory molecule-long); Fas:Fc; FAP-I; NOK2; F2051; F1926;F2928; ZB4; Fas M3 mAb; EGF; 740 Y-P; SC 3036 (KKHTDDGYMPMSPGVA); PI3-kinase Activator (Santa Cruz Biotechnology, Inc.); Pam3Cys((S)-(2,3-bis(palmitoyloxy)-(2RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser(S)-Lys4-OH,trihydrochloride); ActI (NF-kB activator 1); an anti-DcB antibody;Acetyl-11-keto-b-Boswellic Acid; Andrographolide; Caffeic Acid PhenethylEster (CAPE); Gliotoxin; Isohelenin; NEMO-Binding Domain Binding Peptide(DRQIKIWFQNRRMKWKKTALD WSWLQTE); NF-kB Activation Inhibitor(6-Amino-4-(4-phenoxyphenylethylamino)quinazoline); NF-kB ActivationInhibitor II (4-Methyl-N1-(3-phenylpropyl)benzene-1,2-diamine); NF-kBActivation Inhibitor III(3-Chloro-4-nitro-N-(5-nitro-2-thiazolyl)-benzamide); NF-kB ActivationInhibitor IV ((E)-2-Fluoro-4′-methoxystilbene); NF-kB ActivationInhibitor V (5-Hydroxy-(2,6-diisopropylphenyl)-1H-isoindole-1,3-dione);NF-kB SN50 (AAV ALLP A VLLALL AP VQRKRQKLMP); Oridonin; Parthenolide;PPM-18 (2-Benzoylamino-1,4-naphthoquinone); Rol06-9920; Sulfasalazine;TIRAP Inhibitor Peptide (RQIKiWFNRRMKWKKLQLRD AAPGGAIVS); Withaferin A;Wogonin; BAY 11-7082 ((E)3-[(4-Methylphenyl)sulfonyl]-2-propenenitrile);BAY 11-7085 ((E)3-[(4-t-Butylphenyl)sulfonyl]-2-propenenitrile);(E)-Capsaicin; Aurothiomalate (ATM or AuTM); Evodiamine; Hypoestoxide;IKK Inhibitor III (BMS-345541); IKK Inhibitor VII; IKK Inhibitor X; IKKInhibitor II; IKK-2 Inhibitor IV; IKK-2 Inhibitor V; IKK-2 Inhibitor VI;IKK-2 Inhibitor (SC-514); IkB Kinase Inhibitor Peptide; IKK-3 InhibitorIX; ARRY-797 (Array BioPharma); SB-220025(5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinlyl)imidazole);SB-239063(trans-4-[4-(4-Fluorophenyl)-5-(2-methoxy-4-pyrimidinyl)-1H-imidazol-1-yl]cyclohexanol);SB-202190(4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole);JX-401 (-[2-Methoxy-4-(methylthio)benzoyl]-4-(phenylmethyl)piperidine);PD-169316(4-(4-Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-1H-imidazole);SKF-86002(6-(4-Fluorophenyl)-2,3-dihydro-5-(4-pyridinyl)imidazo[2,1-b]thiazoledihydrochloride); SB-200646(N-(I-Methyl-1H-indol-5-yl)-N′-3-pyridinylurea); CMPD-I(2′-Fluoro-N-(4-hydroxyphenyl)-[1,1′-biphenyl]-4-butanamide); EO-1428((2-Methylphenyl)-[4-[(2-amino-4-bromophenyl)amino]-2-chlorophenyl]methanone);SB-253080(4-[5-(4-Fluorophenyl)-2-[4-(methylsulfonyl)phenyl]-1H-imidazol-4-yl]pyridine);SD-169 (1H-Indole-5-carboxamide); SB-203580(4-(4-Fluorophenyl)-2-(4-methylsulfinyl phenyl)-5-(4-pyridyl)1H-imidazole); TZP-101 (Tranzyme Pharma); TZP-102 (Tranzyme Pharma);GHRP-6 (growth hormone-releasing peptide-6); GHRP-2 (growthhormone-releasing peptide-2); EX-1314 (Elixir Pharmaceuticals); MK-677(Merck); L-692,429 (Butanamide,3-amino-3-methyl-N-(2,3,4,5-tetrahydro-2-oxo-1-((2′-(1H-tetrazol-5-yl)(1,1′-biphenyl)-4-yl)methyl)-1H-1-benzazepin-3-yl)-,(R)-); EP1572 (Aib-DTrp-DgTrp-CHO); diltiazem; metabolites of diltiazem;BRE (Brain and Reproductive organ-Expressed protein); verapamil;nimodipine; diltiazem; omega-conotoxin; GVIA; amlodipine; felodipine;lacidipine; mibefradil; NPPB (5-Nitro-2-(3-phenylpropylamino)benzoicAcid); flunarizine; erythropoietin; pipeline; hemin; brazilin; z-VAD-FMK(Benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone); z-LEHD-FMK(benzyloxycarbonyl-Leu-Glu(OMe)-His-Asp(OMe)-fluoromethylketone);B-D-FMK (boc-aspartyl(Ome)-fluoromethylketone); Ac-LEHD-CHO(N-acetyl-Leu-Glu-His-Asp-CHO); Ac-IETD-CHO(N-acetyl-Ile-Glu-Thr-Asp-CHO); z-IETD-FMK(benzyloxycarbonyl-Ile-Glu(OMe)-Thr-Asp(OMe)-fluoromethy Iketone);FAM-LEHD-FMK (benzyloxycarbonyl Leu-Glu-His-Asp-fluoromethyl ketone);FAM-LETD-FMK (benzyloxycarbonyl Leu-Glu-Thr-Asp-fluoromethyl ketone);Q-VD-OPH (Quinoline-Val-ASp-CH2-O-Ph); XIAP; cIAP-1; cIAP-2; ML-IAP;ILP-2; NAIP; Survivin; Bruce; IAPL-3; fortilin; leupeptine; PD-150606(3-(4-Iodophenyl)-2-mercapto-(Z)-2-propenoic acid); MDL-28170(Z-Val-Phe-CHO); calpeptin; acetyl-calpastatin; MG 132(N-[(phenylmethoxy)carbonyl]-L-leucyl-N-[(IS)-1-formyl-3-methylbutyl]-L-leucinamide);MYODUR; BN 82270 (Ipsen); BN 2204 (Ipsen); AHLi-11 (QuarkPharmaceuticals), an mdm2 protein, pifithrin-α(1-(4-Methylphenyl)-2-(4,5,6,7-tetrahydro-2-imino-3(2H)-benzothiazolyl)ethanone);trans-stilbene; cis-stilbene; resveratrol; piceatannol; rhapontin;deoxyrhapontin; butein; chalcon; isoliquirtigen; butein;4,2′,4′-trihydroxychalcone; 3,4,2′,4′,6′-pentahydroxychalcone; flavone;morin; fisetin; luteolin; quercetin; kaempferol; apigenin; gossypetin;myricetin; 6-hydroxyapigenin; 5-hydroxyflavone;5,7,3′,4′,5′-pentahydroxyflavone; 3,7,3′,4′,5′-pentahydroxyflavone;3,6,3′,4′-tetrahydroxyflavone; 7,3′,4′,5′-tetrahydroxyflavone;3,6,2′,4′-tetrahydroxyflavone; 7,4′-dihydroxyflavone;7,8,3′,4′-tetrahydroxy flavone; 3,6,2′, 3′-tetrahydroxyflavone;4′-hydroxyflavone; 5-hydroxyflavone; 5,4′-dihydroxyflavone;5,7-dihydroxyflavone; daidzein; genistein; naringenin; flavanone;3,5,7,3′,4′-pentahydroxyflavanone; pelargonidin chloride; cyanidinchloride; delphinidin chloride; (−)-epicatechin (Hydroxy Sites:3,5,7,3′,4{circumflex over ( )}; (−)-catechin (Hydroxy Sites:3,5,7,3′,40; (−)-gallocatechin (Hydroxy Sites: 3,5,7,3′,4′,50(+)-catechin (Hydroxy Sites: 3,5,7,3′,4{circumflex over ( )};(+)-epicatechin (Hydroxy Sites: 3,5,7,3′,41J; Hinokitiol (b-Thujaplicin;2-hydroxy-4-isopropyl-2,4,6-cycloheptatrien-1-one); L-(+)-Ergothioneine((S)-a-Carboxy-2,3-dihydro-N,N,N-trimethyl-2-thioxo-1H-imidazole4-ethanaminiuminner salt); Caffeic Acid Phenyl Ester; MCI-186(3-Methyl-1-phenyl-2-pyrazolin-5-one); HBED hydroxybenzy{circumflex over( )}ethylenediamine-HN′-diacetic acid«H2O); Ambroxol(trans-4-(2-Amino-3,5-dibromobenzylamino)cyclohexane-HCl; and U-83836E((+2-((4-(2,6-di-1-Pyrrolidinyl-4-pyrimidinyl)-1-piperzainyl)methyl)-3,4-dihydro-2,5,7,8-tetramethyl-2H-1-benzopyran-6-ol*2HCl);/5-1-5-methyl-nicotinamide-2′-deoxyribose;/S-D-1′-5-methyl-nico-tinamide-2′-deoxyribofuranoside;/3-1′-4,5-dimethyl-nicotinamide-2′-de-oxyribose;/3-D-I′-4,5-dimethyl-nicotinamide-2′-deoxyribofuranoside; 1-Naphthyl PP1 (1-(1, 1-Dimethylethyl)-3-(1-naphthalenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine);Lavendustin A(5-[[(2,5-Dihydroxyphenyl)methyl][(2-hydroxyphenyl)methyl]amino]-2-hydroxybenzoicacid); MNS (3,4-Methylenedioxy-b-nitrostyrene); PP 1(1-(1,1-Dimethylethyl)-1-(4-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine);PP2 (3-(4-chlorophenyl)1-(1,1-dimethylethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine); KX1-004(Kinex); KX1-005 (Kinex); KX1-136 (Kinex); KX1-174 (Kinex); KX1-141(Kinex); KX2-328 (Kinex); KXI-306 (Kinex); KX1-329 (Kinex); KX2-391(Kinex); KX2-377 (Kinex); ZD4190 (Astra Zeneca;N-(4-bromo-2-fluorophenyl)-6-methoxy-7-(2-(1H-1,2,3-triazol-1-yl)ethoxy)quinazolin-4-amine);AP22408 (Ariad Pharmaceuticals); AP23236 (Ariad Pharmaceuticals);AP23451 (Ariad Pharmaceuticals); AP23464 (Ariad Pharmaceuticals);AZD0530 (Astra Zeneca); AZM475271 (M475271; Astra Zeneca); Dasatinib(N-(2-chloro-6-methylphneyl)-2-(6-(4-(2-hydroxyethyl)-piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide); GN963(trans-4-(6,7-dimethoxyquinoxalin-2ylamino)cyclohexanol sulfate);Bosutinib(4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-(3-(4-methyl-1-piperazinyl)propoxy)-3-quinolinecarbonitrile);or combinations thereof.

VII. Fluorescent Moieties

In some embodiments, the human neuron or nerve targeting moleculefurther comprises a fluorescent moiety (e.g., a fluorescent protein,peptide, or fluorescent dye molecule). All fluorescent moieties areencompassed within the term “fluorescent moiety.” Specific examples offluorescent moieties given herein, are illustrative and are not meant tolimit the fluorescent moieties for use with the targeting moleculesdisclosed herein. In some embodiments, the human neuron or nervetargeting molecule further comprises a cargo. In some embodiments, thehuman neuron or nerve targeting molecule comprises a peptide sequenceselected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP403; SEQ ID NO:3), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3). In some embodiments, the targeting molecule comprises apeptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2; SEQ ID NO:11),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2; SEQ ID NO:7), SGQVPWEEPYYVVKK(HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20),QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In some embodiments, thetargeting molecule comprises the peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2). In some embodiments, the targeting molecule comprises thepeptide DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments, thetargeting molecule comprises the peptide Ac-SGQVPWEEPYYVVKKSSGGC (HNP401with GGC linker; SEQ ID NO:4). In some embodiments, the targetingmolecule comprises the peptide Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGClinker; SEQ ID NO:5). In some embodiments, the targeting moleculecomprises the peptide Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQID NO:6). In some embodiments, the targeting molecule comprises thepeptide QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments,the targeting molecule comprises the peptide QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21). In some embodiments, thetargeting molecule comprises the peptide Ac-QVPWEEPYYVVKKSSGGC(HNP401-N-2 with GGC linker; SEQ ID NO:7). In some embodiments, thetargeting molecule comprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQID NO:22). In some embodiments, the targeting molecule comprises thepeptide Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).In some embodiments, the targeting molecule comprises the peptideEEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23). In some embodiments, thetargeting molecule comprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6with GGC linker; SEQ ID NO:9). In some embodiments, the targetingmolecule comprises the peptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). Insome embodiments, the targeting molecule comprises the peptideAc-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10). In someembodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some embodiments, thetargeting molecule comprises the peptide Ac-SGQVPWEEPYYVVKKGGC(HNP401-C-2 with GGC linker; SEQ ID NO:11). In some embodiments, thetargeting molecule comprises the peptide SGQVPWEEPYYVV (HNP401-C-4; SEQID NO:26). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27). In some embodiments, thetargeting molecule comprises the peptide Ac-SGQVPWEEPYYGGC (HNP401-C-6with GGC linker; SEQ ID NO:13). In some embodiments, the targetingmolecule comprises the peptide SGQVPWEEP (HNP401-C-8; SEQ ID NO:28). Insome embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14). In someembodiments, the targeting molecule comprises the peptideDTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16). In some embodiments, thetargeting molecule comprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments, thetargeting molecule comprises a peptide that is not Ac-SHSNTQTLAKAPEHTGC(Ac-NP41 with GC linker; SEQ ID NO:17). In some embodiments, thetargeting molecule comprises a peptide that is not SHSNTQTLAKAPEHTGC(NP41 with GC linker; SEQ ID NO:18). In some embodiments, the peptide isnot NTQTLAKAPEHT (NP41; SEQ ID NO:19).

In some embodiments, the peptide or aptamer is directly bound to afluorescent moiety. In some embodiments, the peptide or aptamer isindirectly (e.g., via a linker) bound to a fluorescent moiety. In someembodiments, the peptide or aptamer is bound to a fluorescent moiety atits N-terminus, at its C-terminus, or at an internal position (e.g., toan internal amino acid) of the peptide or aptamer. In some embodiments,two or more peptides or aptamers are directly or indirectly bound to asingle fluorescent moiety.

Examples of fluorescent dyes include, but are not limited to, xanthenes(e.g., rhodamines, rhodols and fluoresceins, and their derivatives);bimanes; coumarins and their derivatives (e.g., umbelliferone andaminomethyl coumarins); aromatic amines (e.g., dansyl; squarate dyes);benzofurans; fluorescent cyanines; carbazoles; dicyanomethylene pyranes;polymethine; oxabenzanthrane; xanthene; pyrylium; carbostyl; perylene;acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene;pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metalchelate complexes; rare-earth metal chelate complexes; and derivativesof such dyes.

In some embodiments, the fluorescent moiety is a fluorescein dye.Examples of fluorescein dyes include, but are not limited to,5-carboxyfluorescein, fluorescein-5-isothiocyanate and6-carboxyfluorescein, 5,6-dicarboxyfluorescein, 5-(and6)-sulfofluorescein, sulfonefluorescein, succinyl fluorescein, 5-(and6)-carboxy SNARF-1, carboxyfluorescein sulfonate, carboxyfluoresceinzwitterion, carbxoyfluorescein quaternary ammonium, carboxyfluoresceinphosphonate, carboxyfluorescein GABA, 5′(6′)-carboxyfluorescein,carboxyfluorescein-cys-Cy5, and fluorescein glutathione.

In some embodiments, the fluorescent moiety is a rhodamine dye. Examplesof rhodamine dyes include, but are not limited to,tetramethylrhodamine-6-isothiocyanate, 5-carboxytetramethylrhodamine,5-carboxy rhodol derivatives, carboxy rhodamine 110, tetramethyl andtetraethyl rhodamine, diphenyldimethyl and diphenyldiethyl rhodamine,dinaphthyl rhodamine, rhodamine 101 sulfonyl chloride (sold under thetradename of TEXAS RED®).

In some embodiments, the fluorescent moiety is a cyanine dye. Examplesof cyanine dyes include, but are not limited to, Cy3, Cy3B, Cy3.5, Cy5,Cy5.5, Cy 7.

In some embodiments, the fluorescent moiety is a peptide. In someembodiments, the fluorescent moiety is Green Fluorescent Protein (GFP).In some embodiments, the fluorescent moiety is a derivative of GFP(e.g., EBFP, EBFP2, Azurite, mKalamal, ECFP, Cerulean, CyPet, YFP,Citrine, Venus, YPet).

Fluorescent labels are detected by any suitable method. For example, afluorescent label may be detected by exciting the fluorochrome with theappropriate wavelength of light and detecting the resultingfluorescence, e.g., by microscopy, visual inspection, via photographicfilm, by the use of electronic detectors such as charge coupled devices(CCDs), photomultipliers, etc.

In some embodiments, the fluorescent moiety is conjugated to highmolecular weight molecule, such as water soluble polymers including, butnot limited to, dextran, PEG, serum albumin, or poly(amidoamine)dendrimer.

Exemplary targeting molecules according to the present inventioninclude: 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ IDNO:104), Ac-SGQVPWEEPYYVVKKSSGGC-5FAM (HNP401 with GGC linker; SEQ IDNO:105), Ac-WEYHYVDLNWTSQHPQGGC-5FAM (HNP402 with GGC linker; SEQ IDNO:106), Ac-DLPDIIWDFNWETAGGC-5FAM (HNP403 with GGC linker; SEQ IDNO:107), Ac-QVPWEEPYYVVKKSSGGC-5FAM (HNP401-N-2 with GGC linker; SEQ IDNO:108), Ac-PWEEPYYVVKKSSGGC-5FAM (HNP401-N-4 with GGC linker; SEQ IDNO:109), Ac-EEPYYVVKKSSGGC-5FAM (HNP401-N-6 with GGC linker; SEQ IDNO:110), Ac-PYYVVKKSSGGC-5FAM (HNP401-N-8 with GGC linker; SEQ IDNO:111), Ac-SGQVPWEEPYYVVKKGGC-5FAM (HNP401-C-2 with GGC linker; SEQ IDNO:112), Ac-SGQVPWEEPYYVVGGC-5FAM (HNP401-C-4 with GGC linker; SEQ IDNO:113), Ac-SGQVPWEEPYYGGC-5FAM (HNP401-C-6 with GGC linker; SEQ IDNO:114), and Ac-SGQVPWEEPGGC-5FAM (HNP401-C-8 with GGC linker; SEQ IDNO:115).

VIII. Linkers

In some embodiments, a cargo (e.g., a fluorescent moiety,photosensitizing agent, or drug) is directly attached to the humanneuron or nerve targeting molecule, e.g. at the end of the targetingpeptide. Alternatively, in some embodiments, a cargo (e.g., afluorescent moiety or drug) is indirectly attached to a targetingmolecule disclosed herein (e.g., via a linker). In some embodiments, thehuman neuron or nerve targeting molecule further comprises a cargo. Insome embodiments, the human neuron or nerve targeting molecule comprisesa peptide sequence selected from the group consisting of:SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP40 with GGC linker 3; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ IDNO:104). In some embodiments, the targeting molecule comprises a peptideselected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ IDNO:11), and Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ IDNO:7). In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In some embodiments, thetargeting molecule comprises the peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2). In some embodiments, the targeting molecule comprises thepeptide DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments, thetargeting molecule comprises the peptide Ac-SGQVPWEEPYYVVKKSSGGC (HNP401with GGC linker; SEQ ID NO:4). In some embodiments, the targetingmolecule comprises the peptide Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGClinker; SEQ ID NO:5). In some embodiments, the targeting moleculecomprises the peptide Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQID NO:6). In some embodiments, the targeting molecule comprises thepeptide QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments,the targeting molecule comprises the peptide QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21). In some embodiments, thetargeting molecule comprises the peptide Ac-QVPWEEPYYVVKKSSGGC(HNP401-N-2 with GGC linker; SEQ ID NO:7). In some embodiments, thetargeting molecule comprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQID NO:22). In some embodiments, the targeting molecule comprises thepeptide Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).In some embodiments, the targeting molecule comprises the peptideEEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23). In some embodiments, thetargeting molecule comprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6with GGC linker; SEQ ID NO:9). In some embodiments, the targetingmolecule comprises the peptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). Insome embodiments, the targeting molecule comprises the peptideAc-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10). In someembodiments, the targeting molecule comprises the peptideSGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some embodiments, thetargeting molecule comprises the peptide Ac-SGQVPWEEPYYVVKKGGC(HNP401-C-2 with GGC linker; SEQ ID NO:11). In some embodiments, thetargeting molecule comprises the peptide SGQVPWEEPYYVV (HNP401-C-4; SEQID NO:26). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).In some embodiments, the targeting molecule comprises the peptideSGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27). In some embodiments, thetargeting molecule comprises the peptide Ac-SGQVPWEEPYYGGC (HNP401-C-6with GGC linker; SEQ ID NO:13). In some embodiments, the targetingmolecule comprises the peptide SGQVPWEEP (HNP401-C-8; SEQ ID NO:28). Insome embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14). In someembodiments, the targeting molecule comprises the peptideDTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16). In some embodiments, thetargeting molecule comprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments, thetargeting molecule comprises a peptide that is not Ac-SHSNTQTLAKAPEHTGC(Ac-NP41 with GC linker; SEQ ID NO:17). In some embodiments, thetargeting molecule comprises a peptide that is not SHSNTQTLAKAPEHTGC(NP41 with GC linker; SEQ ID NO:18). In some embodiments, the targetingmolecule comprises a peptide that is not NTQTLAKAPEHT (SEQ ID NO:19).

As used herein, a “linker” is any molecule capable of binding (e.g.,covalently) to a targeting molecule disclosed herein. Linkers include,but are not limited to, straight or branched-chain carbon linkers,heterocyclic carbon linkers, amino acid linkers (e.g., D- or L-aminoacid), lipophilic residues, peptide linkers, peptide nucleic acidlinkers, hydrazone linkers, SPDB disulfide, sulfo-SPDB, maleimidomethylcyclohexane-1-carboxylate (MCC), aminohexanoic acid linkers, andpolyether linkers (e.g., PEG). For example, poly(ethylene glycol)linkers are available from Quanta Biodesign, Powell, Ohio. These linkersoptionally have amide linkages, sulfhydryl linkages, or heterofunctional linkages.

In some embodiments, the linker binds to a targeting molecule disclosedherein by a covalent linkage. In some embodiments, the covalent linkagecomprises an ether bond, thioether bond, amine bond, amide bond,carbon-carbon bond, carbon-nitrogen bond, carbon-oxygen bond, orcarbon-sulfur bond.

In some embodiments, the linker is flexible. In some embodiments, thelinker is rigid.

In some embodiments, the linker comprises a linear structure. In someembodiments, the linker comprises a non-linear structure. In someembodiments, the linker comprises a branched structure. In someembodiments, the linker comprises a cyclic structure.

In some embodiments, the linker is an alkyl. In some embodiments, thelinker is heteroalkyl.

In some embodiments, the linker is an alkylene. In some embodiments, thelinker is an alkenylene. In some embodiments, the linker is analkynylene. In some embodiments, the linker is a heteroalkylene.

An “alkyl” group refers to an aliphatic hydrocarbon group. The alkylmoiety may be a saturated alkyl or an unsaturated alkyl. Depending onthe structure, an alkyl group can be a monoradical or a diradical (i.e.,an alkylene group).

The“alkyl” moiety may have 1 to 10 carbon atoms (whenever it appearsherein, a numerical range such as “1 to 10” refers to each integer inthe given range; e.g., “1 to 10 carbon atoms” means that the alkyl groupmay consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., upto and including 10 carbon atoms, although the present definition alsocovers the occurrence of the term “alkyl” where no numerical range isdesignated). The alkyl group could also be a “lower alkyl” having 1 to 6carbon atoms. The alkyl group of the compounds described herein may bedesignated as “Ci-C4 alkyl” or similar designations. By way of exampleonly, “C1-C4 alkyl” indicates that there are one to four carbon atoms inthe alkyl chain, i.e., the alkyl chain is selected from methyl, ethyl,propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, and t-butyl. Typicalalkyl groups include, but are in no way limited to, methyl, ethyl,propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl,ethenyl, propenyl, butenyl, and the like.

In some embodiments, the linker comprises a ring structure (e.g., anaryl). As used herein, the term “ring” refers to any covalently closedstructure. Rings include, for example, carbocycles (e.g., aryls andcycloalkyls), heterocycles (e.g., heteroaryls and non-aromaticheterocycles), aromatics (e.g. aryls and heteroaryls), and non-aromatics(e.g., cycloalkyls and non-aromatic heterocycles). Rings can beoptionally substituted. Rings can be monocyclic or polycyclic.

As used herein, the term “aryl” refers to an aromatic ring wherein eachof the atoms forming the ring is a carbon atom. Aryl rings can be formedby five, six, seven, eight, nine, or more than nine carbon atoms. Arylgroups can be optionally substituted. Examples of aryl groups include,but are not limited to phenyl, naphthalenyl, phenanthrenyl, anthracenyl,fluorenyl, and indenyl. Depending on the structure, an aryl group can bea monoradical or a diradical (i.e., an arylene group).

The term “cycloalkyl” refers to a monocyclic or polycyclic non-aromaticradical, wherein each of the atoms forming the ring (i.e. skeletalatoms) is a carbon atom. Cycloalkyls may be saturated, or partiallyunsaturated. Cycloalkyl groups include groups having from 3 to 10 ringatoms. Cycloalkyls include, but are not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

In some embodiments, the ring is a cycloalkane. In some embodiments, thering is a cycloalkene.

In some embodiments, the ring is an aromatic ring. The term “aromatic”refers to a planar ring having a delocalized π-electron systemcontaining 4n+2 π electrons, where n is an integer. Aromatic rings canbe formed from five, six, seven, eight, nine, or more than nine atoms.Aromatics can be optionally substituted. The term “aromatic” includesboth carbocyclic aryl (e.g., phenyl) and heterocyclic aryl (or“heteroaryl” or “heteroaromatic”) groups (e.g., pyridine). The termincludes monocyclic or fused-ring polycyclic (i.e., rings which shareadjacent pairs of carbon atoms) groups.

In some embodiments, the ring is a heterocycle. The term “heterocycle”refers to heteroaromatic and heteroalicyclic groups containing one tofour heteroatoms each selected from O, S and N, wherein eachheterocyclic group has from 4 to 10 atoms in its ring system, and withthe proviso that the ring of said group does not contain two adjacent Oor S atoms. Non-aromatic heterocyclic groups include groups having only3 atoms in their ring system, but aromatic heterocyclic groups must haveat least 5 atoms in their ring system. The heterocyclic groups includebenzo-fused ring systems. An example of a 3-membered heterocyclic groupis aziridinyl. An example of a 4-membered heterocyclic group isazetidinyl (derived from azetidine). An example of a 5-memberedheterocyclic group is thiazolyl. An example of a 6-membered heterocyclicgroup is pyridyl, and an example of a 10-membered heterocyclic group isquinolinyl. Examples of non-aromatic heterocyclic groups arepyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl,tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino,morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl,oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl,diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl,3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl,1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl,dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl,imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl,3H-indolyl and quinolizinyl. Examples of aromatic heterocyclic groupsare pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl,tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl,isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl,benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl,phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl,oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl,naphthyridinyl, and faropyridinyl. The foregoing groups, may beC-attached or N-attached where such is possible. For instance, a groupderived from pyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl(C-attached). Further, a group derived from imidazole may beimidazol-1-yl or imidazol-3-yl (both N-attached) or imidazol-2-yl,imidazol-4-yl or imidazol-5-yl (all C-attached). The heterocyclic groupsinclude benzo-fused ring systems and ring systems substituted with oneor two oxo (═O) moieties such as pyrrolidin-2-one. Depending on thestructure, a heterocycle group can be a monoradical or a diradical(i.e., a heterocyclene group).

In some embodiments, the ring is fused. The term “fused” refers tostructures in which two or more rings share one or more bonds, hi someembodiments, the ring is a dimer. In some embodiments, the ring is atrimer. In some embodiments, the ring is a substituted.

The term “carbocyclic” or “carbocycle” refers to a ring wherein each ofthe atoms forming the ring is a carbon atom. Carbocycle includes aryland cycloalkyl. The term thus distinguishes carbocycle from heterocycle(“heterocyclic”) in which the ring backbone contains at least one atomwhich is different from carbon (i.e., a heteroatom). Heterocycleincludes heteroaryl and heterocycloalkyl. Carbocycles and heterocyclescan be optionally substituted.

In some embodiments, the linker is substituted. The term “optionallysubstituted” or “substituted” means that the referenced group may besubstituted with one or more additional group(s) individually andindependently selected from Ci-Cealkyl, C3-Cgcycloalkyl, aryl,heteroaryl, C2-C6heteroalicyclic, hydroxy, Ci-C6alkoxy, aryloxy,Ci-C6alkylthio, arylthio, Ci-C6alkylsulfoxide, arylsulfoxide,Ci-C6alkylsulfone, arylsulfone, cyano, halo, C2-C8acyl, C2-C8acyloxy,nitro, Ci-C6haloalkyl, Ci-C6fluoroalkyl, and amino, includingCi-C6alkylamino, and the protected derivatives thereof. By way ofexample, an optional substituents may be LSRS, wherein each Ls isindependently selected from a bond, —O—, —C(═O)—, —S—, —S(═O)—, —S(O)2-,—NH—, —NHC(O)—, —C(O)NH—, S(O)2NH—, —NHS(O)2-, —OC(O)NH—, —NHC(O)O—,—(CpC6alkyl)-, or —(C2-C6alkenyl)-; and each Rs is independentlyselected from H, (Ci-C4alkyl), (C3-C8cycloalkyl), heteroaryl, aryl, andCi-C6heteroalkyl. Optionally substituted non-aromatic groups may besubstituted with one or more oxo (═O). The protecting groups that mayform the protective derivatives of the above substituents are known tothose of skill in the art.

In some embodiments, a bifunctional linker having one functional groupreactive with a group on one molecule (e.g., a targeting molecule), andanother group reactive on the other molecule (e.g., a fluorescent moietyor a drug), is used to form the desired conjugate. Alternatively, insome embodiments, derivatization is performed to provide functionalgroups. Thus, for example, procedures for the generation of freesulfhydryl groups on peptides are also known (See U.S. Pat. No.4,659,839). A linker may alternatively comprise a heterobifunctionalcrosslinker comprising two or more different reactive groups that form aheterocyclic ring that can interact with a targeting molecule. Forexample, a heterobifunctional crosslinker such as cysteine may comprisean amine reactive group and a thiol-reactive group can interact with analdehyde on a derivatized targeting molecule. Additional combinations ofreactive groups suitable for heterobifunctional crosslinkers include,for example, amine- and sulfhydryl reactive groups; carbonyl andsulfhydryl reactive groups; amine and photoreactive groups; sulfhydryland photoreactive groups; carbonyl and photoreactive groups; carboxylateand photoreactive groups; and arginine and photoreactive groups.Examples of a heterobifunctional crosslinker include N-Succinimidyl4-(2-pyridyldithio)butanoate (SPDB) and maleimidomethylcyclohexane-1-carboxylate (MCC).

In some embodiments, a peptide linker consisting of one or more aminoacids is used to join the targeting molecule and a fluorescent moiety ordrug. Generally the peptide linker will have no specific biologicalactivity other than to join the molecules or to preserve some minimumdistance or other spatial relationship between them. However, theconstituent amino acids of the linker may be selected to influence someproperty of the molecule such as the folding, net charge, orhydrophobicity. In some embodiments the peptide linker is relativelyshort, typically less than about 10 amino acids, preferably less thanabout 8 amino acids and more preferably less than 5 amino acids.Non-limiting illustrative examples include glycine and glycine-serinelinkers which can be added to the C-terminus of a targeting peptide. Insome embodiments, a peptide linker is a glycine-glycine-glycine-cysteine(GGGC) linker, a glycine-glycine-cysteine (GGC) linker, aglycine-glycine (GG) linker, or a cysteine (C) linker. In someembodiments, the GGGC, GGC, GG, or C linker is added to the C-terminusof a targeting peptide.

IX. Further Modifications

In some embodiments, the human neuron or nerve targeting molecules ofthe present invention are optionally conjugated to high molecular weightmolecules that increase the multivalency and avidity of labeling. Insome embodiments, the high molecular weight molecules are water-solublepolymers. Examples of suitable water-soluble polymers include, but arenot limited to, peptides, saccharides, poly(vinyls), poly(ethers),poly(amines), poly(carboxylic acids) and the like. In some embodiments,the water-soluble polymers is dextran, polyethylene glycol (PEG),polyoxyalkylene, polysialic acid, starch, or hydroxyethyl starch. Anysuitable method is used to conjugate peptides to water-soluble polymers(see, Hermanson G., Bioconjugate Techniques 2nd Ed., Academic Press,Inc. 2008). In some embodiments, the human neuron or nerve targetingmolecule further comprises a cargo. In some embodiments, the humanneuron or nerve targeting molecule comprises a peptide sequence selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22), EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS(HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ IDNO:104). In some embodiments, the targeting molecule comprises a peptideselected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ IDNO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), Ac-QVPWEEPYYVVKKSS(HNP401-N-2; SEQ ID NO:20), and Ac-QVPWEEPYYVVKKSSGG (HNP401-N-2 with GGlinker; SEQ ID NO:21). In some embodiments, the targeting moleculecomprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In someembodiments, the targeting molecule comprises the peptideWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some embodiments, thetargeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3). In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In someembodiments, the targeting molecule comprises the peptideAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5). In someembodiments, the targeting molecule comprises the peptideAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In someembodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, thetargeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2with GG linker; SEQ ID NO:21). In some embodiments, the targetingmolecule comprises the peptide Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 withGGC linker; SEQ ID NO:7). In some embodiments, the targeting moleculecomprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In someembodiments, the targeting molecule comprises the peptideAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In someembodiments, the targeting molecule comprises the peptide EEPYYVVKKSS(HNP401-N-6; SEQ ID NO:23). In some embodiments, the targeting moleculecomprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQID NO:9). In some embodiments, the targeting molecule comprises thepeptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In some embodiments, thetargeting molecule comprises the peptide Ac-PYYVVKKSSGGC (HNP401-N-8with GGC linker; SEQ ID NO:10). In some embodiments, the targetingmolecule comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ IDNO:25). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ IDNO:11). In some embodiments, the targeting molecule comprises thepeptide SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26). In some embodiments,the targeting molecule comprises the peptide Ac-SGQVPWEEPYYVVGGC(HNP401-C-4 with GGC linker; SEQ ID NO:12). In some embodiments, thetargeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ IDNO:27). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). Insome embodiments, the targeting molecule comprises the peptide SGQVPWEEP(HNP401-C-8; SEQ ID NO:28). In some embodiments, the targeting moleculecomprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQID NO:14). In some embodiments, the targeting molecule comprises thepeptide DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16). In some embodiments,the targeting molecule comprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments, thetargeting molecule comprises a peptide that is not Ac-SHSNTQTLAKAPEHTGC(Ac-NP41 with GC linker; SEQ ID NO:17). In some embodiments, thetargeting molecule comprises a peptide that is not SHSNTQTLAKAPEHTGC(NP41 with GC linker; SEQ ID NO:18). In some embodiments, the targetingmolecule comprises a peptide that is not NTQTLAKAPEHT (SEQ ID NO:19).

In some embodiments, the targeting molecules of the present inventionare conjugated to factors having neurotrophic properties (e.g.,neurotrophic proteins such as nerve growth factor (NGF), brain-derivedneurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4(NT-4), glial cell line-derived neurotrophic factor (GDNF), ciliaryneurotrophic factor (CNTF) as well as non-protein small molecules withneurotrophic properties).

In some embodiments, the targeting molecules of the present inventionare modified to increase solubility. Peptide modifications that increasesolubility include addition of hyphilic amino acid(s), a PEG moiety, orboth. In some embodiments, a PEG moiety is 8-Amino-3,6-dioxaoctanoicacid (AEEA); 12-amino-4,7,10-trioxadodecanoic acid; or15-amino-4,7,10,13-tetraoxapenta-decanoic acid. In some embodiments,about one to ten (e.g., one, two, three, four, five, six, seven, eight,nine, or ten) hydrophilic amino acids may be added to the N-terminus,C-terminus, an internal position, or any combination thereof, of thetargeting molecule to increase solubility. Hydrophilic amino acidsinclude D, E, H, K, N, Q, R, S, T, and G. In some embodiments, thetargeting molecule comprises a K, KK, G, or GG at the N-terminus orC-terminus.

X. Multidomain Targeting Molecules

In certain embodiments, the human neuron or nerve targeting moleculesprovided herein are multidomain neuron or nerve targeting moleculescomprising two or more neuron or nerve targeting peptides, wherein thefirst peptide comprises SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), or5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the first peptide comprises SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), orDLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments, the firstpeptide comprises SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS (HNP401-N-2;SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ IDNO:21), or 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ IDNO:104). In some embodiments, the first peptide comprisesSGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In some embodiments, the firstpeptide comprises WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In someembodiments, the first peptide comprises DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3). In some embodiments, the first peptide comprisesAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In someembodiments, the first peptide comprises Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5). In some embodiments, the first peptidecomprises Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). Insome embodiments, the first peptide comprises QVPWEEPYYVVKKSS(HNP401-N-2; SEQ ID NO:20). In some embodiments, the first peptidecomprises QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).In some embodiments, the first peptide comprises Ac-QVPWEEPYYVVKKSSGGC(HNP401-N-2 with GGC linker; SEQ ID NO:7). In some embodiments, thefirst peptide comprises PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). Insome embodiments, the first peptide comprises Ac-PWEEPYYVVKKSSGGC(HNP401-N-4 with GGC linker; SEQ ID NO:8). In some embodiments, thefirst peptide comprises EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23). In someembodiments, the first peptide comprises Ac-EEPYYVVKKSSGGC (HNP401-N-6with GGC linker; SEQ ID NO:9). In some embodiments, the first peptidecomprises PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In some embodiments, thefirst peptide comprises Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQID NO:10). In some embodiments, the first peptide comprisesSGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some embodiments, thefirst peptide comprises Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGClinker; SEQ ID NO:11). In some embodiments, the first peptide comprisesSGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26). In some embodiments, the firstpeptide comprises Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQID NO:12). In some embodiments, the first peptide comprises SGQVPWEEPYY(HNP401-C-6; SEQ ID NO:27). In some embodiments, the first peptidecomprises Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).In some embodiments, the first peptide comprises SGQVPWEEP (HNP401-C-8;SEQ ID NO:28). In some embodiments, the first peptide comprisesAc-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14). In someembodiments, the first peptide comprises DTHAHAKPRVPAFKSV (HNP 404; SEQID NO:16). In some embodiments, the first peptide comprises5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

The two or more neuron or nerve targeting peptides within a multidomaintargeting molecule can be the same neuron or nerve targeting peptide, orare preferably different neuron or nerve targeting peptides. In someembodiments, multidomain targeting molecules comprise a second peptidecomprising: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ(HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3),DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104), SHSSEFPRSWDMETN (HNP301; SEQID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30); SHSTMKTLSL (HNP305; SEQ IDNO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ IDNO:33), HKTAQWPFIAFR (NP123; SEQ ID NO:34), RLTNAPAYQAPA (NP124; SEQ IDNO:35), MQNPLNGKPGR (NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ IDNO:37), YPSPNRPPNLTN (NP127; SEQ ID NO:38), or NTQTLAKAPEHTG (NP117; SEQID NO:39).

In some embodiments, the first neuron or nerve targeting peptide isselected from the group consisting of: QVPWEEPYYVVKKSS (HNP401-N-2; SEQID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21),SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments, the second peptide is selected from the groupconsisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104), SHSSEFPRSWDMETN (HNP301; SEQID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30); SHSTMKTLSL (HNP305; SEQ IDNO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ IDNO:33), HKTAQWPFIAFR (NP123; SEQ ID NO:34), RLTNAPAYQAPA (NP124; SEQ IDNO:35), MQNPLNGKPGR (NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ IDNO:37), YPSPNRPPNLTN (NP127; SEQ ID NO:38), or NTQTLAKAPEHTG (NP117; SEQID NO:39).

In some embodiments, the first neuron or nerve targeting peptidecomprises QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In someembodiments, the first neuron or nerve targeting peptide isQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, thesecond peptide is selected from the group consisting of:SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), and SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), SHSSEFPRSWDMETN (HNP301; SEQID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30); SHSTMKTLSL (HNP305; SEQ IDNO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ IDNO:33), HKTAQWPFIAFR (NP123; SEQ ID NO:34), RLTNAPAYQAPA (NP124; SEQ IDNO:35), MQNPLNGKPGR (NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ IDNO:37), YPSPNRPPNLTN (NP127; SEQ ID NO:38), or NTQTLAKAPEHTG (NP117; SEQID NO:39).

In some embodiments, the first neuron or nerve targeting peptidecomprises QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).In some embodiments, the first neuron or nerve targeting peptide isQVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21). In someembodiments, the second peptide is selected from the group consistingof: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402;SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV(HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGClinker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker;SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ IDNO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104), SHSSEFPRSWDMETN (HNP301; SEQID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30); SHSTMKTLSL (HNP305; SEQ IDNO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ IDNO:33), HKTAQWPFIAFR (NP123; SEQ ID NO:34), RLTNAPAYQAPA (NP124; SEQ IDNO:35), MQNPLNGKPGR (NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ IDNO:37), YPSPNRPPNLTN (NP127; SEQ ID NO:38), or NTQTLAKAPEHTG (NP117; SEQID NO:39).

In some embodiments, the first neuron or nerve targeting peptidecomprises SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In someembodiments, the first neuron or nerve targeting peptide isSGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some embodiments, thesecond peptide is selected from the group consisting of:SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104), SHSSEFPRSWDMETN (HNP301; SEQID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30); SHSTMKTLSL (HNP305; SEQ IDNO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ IDNO:33), HKTAQWPFIAFR (NP123; SEQ ID NO:34), RLTNAPAYQAPA (NP124; SEQ IDNO:35), MQNPLNGKPGR (NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ IDNO:37), YPSPNRPPNLTN (NP127; SEQ ID NO:38), or NTQTLAKAPEHTG (NP117; SEQID NO:39).

In some embodiments, the first neuron or nerve targeting peptidecomprises 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ IDNO:104). In some embodiments, the first neuron or nerve targetingpeptide is 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ IDNO:104). In some embodiments, the second peptide is selected from thegroup consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2(HNP401-N-2 with GG linker; SEQ ID NO:104), SHSSEFPRSWDMETN (HNP301; SEQID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30); SHSTMKTLSL (HNP305; SEQ IDNO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ IDNO:33), HKTAQWPFIAFR (NP123; SEQ ID NO:34), RLTNAPAYQAPA (NP124; SEQ IDNO:35), MQNPLNGKPGR (NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ IDNO:37), YPSPNRPPNLTN (NP127; SEQ ID NO:38), or NTQTLAKAPEHTG (NP117; SEQID NO:39).

In some embodiments, the neuron or nerve targeting peptides within amultidomain targeting molecule are directly bound to each other. In someembodiments, the neuron or nerve targeting peptides within a multidomaintargeting molecule are indirectly bound to each other, e.g., via alinker or cargo. In some embodiments, the targeting peptides arearranged in a linear fashion. In some embodiments, the targetingpeptides of a multidomain targeting molecule are arranged in a branchedstructure. In some embodiments, a multidomain targeting moleculecomprises two, three, four, five, or more neuron or nerve targetingpeptides.

XI. Methods of Labeling

Disclosed herein, in certain embodiments, are methods of labeling aneuron or nerve (or component of either) by contacting a neuron or nervewith a human neuron or nerve targeting molecule described herein. Insome embodiments, the human neuron or nerve targeting molecule furthercomprises a cargo. In some embodiments, the human neuron or nervetargeting molecule comprises a peptide sequence selected from the groupconsisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22), EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS(HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3). In some embodiments, the targeting molecule comprises apeptide selected from the group consisting of SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker;SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ IDNO:7), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS(HNP401-N-2; SEQ ID NO:20), and QVPWEEPYYVVKKSSGG (HNP401-N-2 with GGlinker; SEQ ID NO:21). In some embodiments, the targeting moleculecomprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In someembodiments, the targeting molecule comprises the peptideWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some embodiments, thetargeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3). In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In someembodiments, the targeting molecule comprises the peptideAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5). In someembodiments, the targeting molecule comprises the peptideAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In someembodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, thetargeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2with GG linker; SEQ ID NO:21). In some embodiments, the targetingmolecule comprises the peptide Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 withGGC linker; SEQ ID NO:7). In some embodiments, the targeting moleculecomprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In someembodiments, the targeting molecule comprises the peptideAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In someembodiments, the targeting molecule comprises the peptide EEPYYVVKKSS(HNP401-N-6; SEQ ID NO:23). In some embodiments, the targeting moleculecomprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQID NO:9). In some embodiments, the targeting molecule comprises thepeptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In some embodiments, thetargeting molecule comprises the peptide Ac-PYYVVKKSSGGC (HNP401-N-8with GGC linker; SEQ ID NO:10). In some embodiments, the targetingmolecule comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ IDNO:25). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ IDNO:11). In some embodiments, the targeting molecule comprises thepeptide SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26). In some embodiments,the targeting molecule comprises the peptide Ac-SGQVPWEEPYYVVGGC(HNP401-C-4 with GGC linker; SEQ ID NO:12). In some embodiments, thetargeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ IDNO:27). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). Insome embodiments, the targeting molecule comprises the peptide SGQVPWEEP(HNP401-C-8; SEQ ID NO:28). In some embodiments, the targeting moleculecomprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GGlinker; SEQ ID NO:104). In some embodiments, the targeting moleculecomprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQID NO:14). In some embodiments, the targeting molecule comprises thepeptide DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16). In some embodiments,the targeting molecule comprises a peptide that is notAc-SHSNTQTLAKAPEHTGC (Ac-NP41 with GC linker; SEQ ID NO:17). In someembodiments, the targeting molecule comprises a peptide that is notSHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID NO:18). In someembodiments, the targeting molecule comprises a peptide that is notNTQTLAKAPEHT (SEQ ID NO:19).

In some embodiments, a first human neuron or nerve targeting molecule isadministered in combination (simultaneously, concurrently, or serially)with a second human neuron or nerve targeting molecule. In furtherembodiments, the first targeting molecule, the second targetingmolecule, or both comprise a cargo. In yet further embodiments, thecargo of the first targeting molecule, the cargo of the second targetingmolecule, or both are fluorescent moeities, which may be the samefluorescent moeities or different fluorescent moieties.

In some embodiments, a human neuron or nerve targeting molecule isadminstered in combination (simultaneously, concurrently, or serially)with a fluorescent moiety (e.g., fluorescent moiety is not conjugated tothe targeting molecule, “free” fluorescent moiety). In some embodiments,the fluorescent moiety is a fluorescein, e.g., carboxyfluorescein.

In some embodiments, the contacting occurs in vivo. In some embodiments,the contacting occurs in vitro.

In some embodiments, a neuron or nerve (or component thereof) is labeledfor identification during surgery. In some embodiments the surgery iscancer surgery. In some embodiments the cancer is selected from thegroup consisting of prostate cancer, liver cancer (HCC), colorectalcancer, ovarian cancer, endometrial cancer, breast cancer, pancreaticcancer, stomach cancer, cervical cancer, head and neck cancer, thyroidcancer, testis cancer, urothelial cancer, lung cancer, melanoma,testicular germ cell tumors, mesothelioma, and esophageal cancer. Insome embodiments, the cancer is prostate cancer. In some embodiments,the method comprises administering a targeting molecule disclosed hereinto a subject that will undergo surgery. In some embodiments, the methodcomprises administering a targeting molecule disclosed herein to asubject that is undergoing surgery. In some embodiments, a targetingmolecule disclosed herein is administered to a patient systemically. Insome embodiments, a targeting molecule disclosed herein is administeredto a patient locally.

XII. Drug Delivery

Disclosed herein, in certain embodiments, are methods of targeted drugdelivery. In some embodiments, a human neuron or nerve targetingmolecule disclosed herein delivers a drug to a specific target. In someembodiments, a targeting molecule disclosed herein delivers a drug to aneuron or nerve. In some embodiments, the human neuron or nervetargeting molecule further comprises a cargo. In some embodiments, thehuman neuron or nerve targeting molecule comprises a peptide sequenceselected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16),Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4),Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22), EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS(HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3). In some embodiments, the targeting molecule comprises apeptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker;SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ IDNO:7), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS(HNP401-N-2; SEQ ID NO:20), and QVPWEEPYYVVKKSSGG (HNP401-N-2 with GGlinker; SEQ ID NO:21). In some embodiments, the targeting moleculecomprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In someembodiments, the targeting molecule comprises the peptideWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some embodiments, thetargeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3). In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In someembodiments, the targeting molecule comprises the peptideAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5). In someembodiments, the targeting molecule comprises the peptideAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In someembodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, thetargeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2with GG linker; SEQ ID NO:21). In some embodiments, the targetingmolecule comprises the peptide Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 withGGC linker; SEQ ID NO:7). In some embodiments, the targeting moleculecomprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In someembodiments, the targeting molecule comprises the peptideAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In someembodiments, the targeting molecule comprises the peptide EEPYYVVKKSS(HNP401-N-6; SEQ ID NO:23). In some embodiments, the targeting moleculecomprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQID NO:9). In some embodiments, the targeting molecule comprises thepeptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In some embodiments, thetargeting molecule comprises the peptide Ac-PYYVVKKSSGGC (HNP401-N-8with GGC linker; SEQ ID NO:10). In some embodiments, the targetingmolecule comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ IDNO:25). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ IDNO:11). In some embodiments, the targeting molecule comprises thepeptide SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26). In some embodiments,the targeting molecule comprises the peptide Ac-SGQVPWEEPYYVVGGC(HNP401-C-4 with GGC linker; SEQ ID NO:12). In some embodiments, thetargeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ IDNO:27). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). Insome embodiments, the targeting molecule comprises the peptide SGQVPWEEP(HNP401-C-8; SEQ ID NO:28). In some embodiments, the targeting moleculecomprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GGlinker; SEQ ID NO:104). In some embodiments, the targeting moleculecomprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQID NO:14). In some embodiments, the targeting molecule comprises thepeptide DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16). In some embodiments,the targeting molecule comprises a peptide that is notAc-SHSNTQTLAKAPEHTGC (Ac-NP41 with GC linker; SEQ ID NO:17). In someembodiments, the targeting molecule comprises a peptide that is notSHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID NO:18). In someembodiments, the targeting molecule comprises a peptide that is notNTQTLAKAPEHT (SEQ ID NO:19).

In some embodiments, the drug is an agent that reduces pain (either theperception of pain or activity of a painful stimulant). In someembodiments, the drug is an anesthetic. In some embodiments, the drug isbenzocaine; carticaine; cinchocaine; cyclomethycaine; lidocaine;prilocaine; propxycaine; proparacaine; tetracaine; tocainide; andtrimecaine; or a combination thereof.

In some embodiments, the drug is an agent that modulates death (e.g.,via apoptosis or necrosis) of a neuron or nerve. In some embodiments,the drug is a cytotoxic agent. In some embodiments, the drug ismethotrexate (RHEUMATREX®, Amethopterin); cyclophosphamide(CYTOXAN®);thalidomide (THALIDOMID®); paclitaxel; pemetrexed;pentostatin; pipobroman; pixantrone; plicamycin; procarbazine;proteasome inhibitors (e.g.; bortezomib); raltitrexed; rebeccamycin;rubitecan; SN-38; salinosporamide A; satraplatin; streptozotocin;swainsonine; tariquidar; taxane; tegafur-uracil; temozolomide;testolactone; tbioTEPA; tioguanine; topotecan; trabectedin; tretinoin;triplatin tetranitrate; tris(2-chloroethyl)amine; troxacitabine; uracilmustard; valrubicin; vinblastine; vincristine; vinorelbine; vorinostat;zosuquidar; or a combination thereof. In some embodiments, the drug is apro-apoptotic agent. In some embodiments, the drug is an anti-apoptoticagent. In some embodiments, the drug is selected from minocycline;SB-203580 (4-(4-Fluorophenyl)-2-(4-methylsulfmyl phenyl)-5-(4-pyridyl)1H-imidazole); PD 169316(4-(4-Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-1H-imidazole); SB202190(4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole); RWJ67657(4-[4-(4-fluorophenyl)-1-(3-phenylpropyl)-5-(4-pyridinyl)-1H-imidazol-2-yl]-3-butyn-1-ol);SB 220025(5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinlyl)imidazole);D-JNKI-1 ((D)-hJIP 175_i 57-DPrO-DPrO-(D)-HIV-TAT57-48); AM-111 (Auris);SP600125 (anthra[1,9-cd]pyrazol-6(2H)-one); JNK Inhibitor I ((L)-HIV-TAT48-57-PP-JBD20); JNK Inhibitor III((L)-HIV-TAT47-57-gaba-c-Jun633-57); AS601245 (1,3-benzothiazol-2-yl(2-[[2-(3-pyridinyl) ethyl] amino]-4 pyrimidinyl) acetonitrile); JNKInhibitor VI (H2N-RPKRPTTLNLF-NH2); JNK Inhibitor VIII(N-(4-Amino-5-cyano-6-ethoxypyridin-2-yl)-2-(2,5-dimethoxyphenyl)acetamide);JNK Inhibitor IX(N-(3-Cyano-4,5,6,7-tetrahydro-1-benzothien-2-yl)-1-naphthamide);dicumarol (3,3′-Methylenebis(4-hydroxycoumarin)); SC-236(4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzene-sulfonamide);CEP-1347 (Cephalon); CEP-11004 (Cephalon); an artificial proteincomprising at least a portion of a Bcl-2 polypeptide; a recombinant FNK;V5 (also known as Bax inhibitor peptide V5); Bax channel blocker((±)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-01); Baxinhibiting peptide P5 (also known as Bax inhibitor peptide P5); Kp7-6;FAIM(S) (Fas apoptosis inhibitory molecule-short); FAIM(L) (Fasapoptosis inhibitory molecule-long);Fas: Fc; FAP-1; NOK2; F2051; F1926;F2928; ZB4; Fas M3 mAb; EGF; 740 Y-P; SC 3036 (KKHTDDGYMPMSPGVA); PI3-kinase Activator (Santa Cruz Biotechnology, Inc.); Pam3Cys((S)-(2,3-bis(palmitoyloxy)-(2RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser(S)-Lys4-OH,trihydrochloride); ActI (NF-kB activator 1); an anti-DcB antibody;Acetyl-11-keto-b-Boswellic Acid; Andrographolide; Caffeic Acid PhenethylEster (CAPE); Gliotoxin; Isohelenin; NEMO-Binding Domain Binding Peptide(DRQIKIWFQNRRMKWKKTALDWSWLQTE); NF-kB Activation Inhibitor(6-Amino-4-(4-phenoxyphenylethylamino)quinazoline); NF-kB ActivationInhibitor II (4-Methyl-N1-(3-phenylpropyl)benzene-1,2-diamine); NF-kBActivation Inhibitor III(3-Chloro-4-nitro-N-(5-nitro-2-thiazolyl)-benzamide); NF-kB Activationinhibitor IV ((E)-2-Fluoro-4′-methoxystilbene); NF-kB ActivationInhibitor V (5-Hydroxy-(2,6-diisopropylphenyl)-1H-isoindole-1,3-dione);NF-kB SN50 (AA VALLP A VLLALLAP VQRKRQKLMP); Oridonin; Parthenolide;PPM-18 (2-Benzoylamino-1,4-naphthoquinone); Rol06-9920; Sulfasalazine;TIRAP Inhibitor Peptide (RQIKIWFNRRMKWKKLQLRD AAPGG AIVS); Withaferin A;Wogonin; BAY 11-7082 ((E)3-[(4-Methylphenyl)sulfonyl]-2-propenenitrile);BAY 11-7085 ((E)3-[(4-t-Butylphenyl)sulfonyl]-2-propenenitrile);(E)-Capsaicin; Aurothiomalate (ATM or AuTM); Evodiamine; Hypoestoxide;IKK Inhibitor III (BMS-345541); IKK Inhibitor VII; IKK Inhibitor X; IKKInhibitor II; IKK-2 Inhibitor IV; IKK-2 Inhibitor V; IKK-2 Inhibitor VI;IKK-2 Inhibitor (SC-514); IkB Kinase Inhibitor Peptide; IKK-3 InhibitorLX; ARRY-797 (Array BioPharma); SB-220025(5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinlyl)imidazole);SB-239063(trans-4-[4-(4-Fluorophenyl)-5-(2-methoxy-4-pyrimidinyl)-1H-imidazol-1-yl]cyclohexanol);SB-202190 (4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole); JX-401(-[2--Methoxy-4-(methylthio)benzoyl]-4-(phenylmethyl)piperidine);PD-169316(4-(4-Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-1H-imidazole);SKF-86002(6-(4-Fluorophenyl)-2,3-dihydro-5-(4-pyridinyl)imidazo[2,1-b]thiazoledihydrochloride); SB-200646(N-(1-Methyl-1H-indol-5-yl)-N′-3-pyridinylurea); CMPD-I(2′-Fluoro-N-(4-hydroxyphenyl)-[1,1′-biphenyl]-4-butanamide); EO-1428((2-Methylphenyl)-[4-[(2-amino-4-bromophenyl)amino]-2-chlorophenyl]methanone);SB-253080(4-[5-(4-Fluorophenyl)-2-[4-(methylsulfonyl)phenyl]-1H-imidazol-4-yl]pyridine);SD-169 (1H-Indole-5-carboxamide); SB-203580(4-(4-Fluorophenyl)-2-(4-methylsulfmyl phenyl)-5-(4-pyridyl)1H-imidazole); TZP-101 (Tranzyme Pharma); TZP-102 (Tranzyme Pharma);GHRP-6 (growth hormone-releasing peptide-6); GHRP-2 (growthhormone-releasing peptide-2); EX-1314 (Elixir Pharmaceuticals); MK-677(Merck); L-692,429 (Butanamide,3-amino-3-methyl-N-(2,3,4,5-tetrahydro-2-oxo-1-((2′-(1H-tetrazol-5-yl)(1,1′-biphenyl)-4-yl)methyl)-1H-1-benzazepin-3-yl)-,(R)-); EP1572 (Aib-DTrp-DgTϕ-CHO); diltiazem; metabolites of diltiazem;BRE (Brain and Reproductive organ-Expressed protein); verapamil;nimodipine; diltiazem; omega-conotoxin; GVIA; amlodipine; felodipine;lacidipine; mibefradil; NPPB (5-Nitro-2-(3-phenylpropylamino)benzoicAcid); flunarizine; erythropoietin; piperine; hemin; brazilin; z-VAD-FMK (Benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone);z-LEHD-FMK(benzyloxycarbonyl-Leu-Glu(OMe)-His-Asp(OMe)-fluoromethylketone);B-D-FMK (boc-aspartyl(Ome)-fluoromethylketone); Ac-LEHD-CHO(N-acetyl-Leu-Glu-His-Asp-CHO); Ac-IETD-CHO(N-acetyl-Ile-Glu-Thr-Asp-CHO); z-IETD-FMK(benzyloxycarbonyl-Ile-Glu(OMe)-Thr-Asp(OMe)-fluoromethy Iketone);FAM-LEHD-FMK (benzyloxycarbonyl Leu-Glu-His-Asp-fluoromethyl ketone);FAM-LETD-FMK (benzyloxycarbonyl Leu-Glu-Thr-Asp-fluoromethyl ketone);Q-VD-OPH (Quinoline-Val- ASp-CH2-O-Ph); XIAP; cIAP-1; cIAP-2; ML-IAP;ILP-2; NAIP; Survivin; Brace; IAPL-3; fortilin; leupeptine; PD-150606(3-(4-Iodophenyl)-2-mercapto-(Z)-2-propenoic acid); MDL-28170(Z-Val-Phe-CHO); calpeptin; acetyl-calpastatin; MG 132(N-[(phenylmethoxy)carbonyl]-L-leucyl-N-[(1S)-1-formyl-3-methylbutyl]-L-leucinamide);MYODUR; BN 82270 (Ipsen); BN 2204 (Ipsen); AHLi-11 (QuarkPharmaceuticals), an mdm2 protein, pifithrin-α(1-(4-Methylphenyl)-2-(4,5,6,7-tetrahydro-2-imino-3(2H)-benzothiazolyl)ethanone);trans-stilbene; cis-stilbene; resveratrol; piceatannol; rhapontin;deoxyrhapontin; butein; chalcon; isoliquirtigen; butein;4,2′,4′-trihydroxychalcone; 3,4,2′,4′,6′-pentahydroxychalcone; flavone;morin; fisetin; luteolin; quercetin; kaempferol; apigenin; gossypetin;myricetin; 6-hydroxyapigenin; 5-hydroxyflavone; 5,7,3′,4′,5′-pentahydroxyflavone; 3,7,3′,4′,5′-pentahydroxyflavone;3,6,3′,4′-tetrahydroxyflavone; 7,3′,4′,5′-tetrahydroxyflavone;3,6,2′,4′-tetrahydroxyflavone; 7,4′-dihydroxyflavone;7,8,3′,4′-tetrahydroxyflavone; 3, 6,2′,3′tetrahydroxyflavone;4′-hydroxyflavone; 5-hydroxyflavone; 5,4′-dihydroxyflavone;5,7-dihydroxyflavone; daidzein; genistein; naringenin; flavanone;3,5,7,3′,4′-pentahydroxyflavanone; pelargonidin chloride; cyanidinchloride; delphinidin chloride; (−)-epicatechin (Hydroxy Sites:3,5,7,3′,4{circumflex over ( )}; (−)-catechin (Hydroxy Sites:3,5,7,3′,4); (−)-gallocatechin (Hydroxy Sites: 3,5,7,3′,4′,5)(+)-catechin (Hydroxy Sites: 3,5,7,3′,4{circumflex over ( )};(+)-epicatechin (Hydroxy Sites: 3,5,7,3′,40; Hinokitiol (b-Thujaplicin;2-hydroxy-4-isopropyl-2,4,6-cycloheptatrien-1-one); L-(+)-Ergothioneine((S)-a-Carboxy-2,3-dihydro-N,N,N-trimethyl-2-thioxo-1H-imidazole4-ethanaminium inner salt); Caffeic Acid Phenyl Ester; MCI-186(3-Methyl-1-phenyl-2-pyrazolin-5-one); HBED(N,N′-Di-(2-hydroxybenzyl)ethylenediamine-N,N′-diacetic acid*H2O);Ambroxol (trans-4-(2-Amino-3,5-dibromobenzylamino)cyclohexane-HCl; andU-83836E((−)-2-((4-(2,6-di-1-Pyrrolidinyl-4-pyrimidinyl)-1-piperzainyl)methyl)-3,4-dihydro-2,5,7,8-tetramethyl-2H-1-benzopyran-6-ol»2HCl);β-1-5-methyl-nicotinamide-2′-deoxyribose;/3-D-1′-5-methyl-nico-tinamide-2′-deoxyribofuranoside;/3-1′-4,5-dimethyl-nicotinamide-2′-de-oxyribose;/3-D-1′-4,5-dimethyl-nicotmamide-2′-deoxyribofuranoside; 1-NaphthylPP1(1-(1,1-Dimethylethyl)-3-(1-naphthalenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine);Lavendustin A (5-[[(2,5-Dihydroxyphenyl)methyl][(2-hydroxyphenyl)methy1] amino]-2-hydroxybenzoic acid); MNS(3,4-Methylenedioxy-b-nitrostyrene); PPI(1-(1,1-Dimethylethyl)-1-(4-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine);PP2(3-(4-chlorophenyl)1-(1,1-dimethylethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine);KX1-004 (Kinex); KX1-005 (Kinex); KX1-136 (Kinex); KX1-174 (Kinex);KX1-141 (Kinex); KX2-328 (Kinex); KX1-306 (Kinex); KX1-329 (Kinex);KX2-391 (Kinex); KX2-377 (Kinex); ZD4190 (Astra Zeneca;N-(4-bromo-2-fluorophenyl)-6-methoxy-7-(2-(1H-1,2,3-triazol-1-yl)ethoxy)quinazolin-4-amine);AP22408 (Ariad Pharmaceuticals); AP23236 (Ariad Pharmaceuticals);AP23451 (Ariad Pharmaceuticals); AP23464 (Ariad Pharmaceuticals);AZD0530 (Astra Zeneca); AZM475271 (M475271; Astra Zeneca); Dasatinib(N-(2-chloro-6-methylphneyl)-2-(6-(4-(2-hydroxyethyl)-piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide); GN963(trans-4-(6,7-dimethoxyquinoxalin-2ylamino)cyclohexanol sulfate);Bosutinib(4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-(3-(4-methyl-1-piperazinyl)propoxy)-3-quinolinecarbonitrile);or combinations thereof.

In some embodiments, the drug is an agent that reduces undesired neuronor nerve impulses. In some embodiments, the drug reduces one or moresymptoms of dyskinesia or synkinesia. In some embodiments, the drug iscarbamazepine, oxcarbazepine, phenytein, valproic acid, sodiumvalproate, cinnarizine, flunarizine, or nimodipine, or combinationsthereof.

In some embodiments, the drug is an agent that promotes regeneration ofneuron or nerve tissue. In some embodiments, the drug is a growthfactor. In some embodiments, the drug is selected from brain-derivedneurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glialcell-line derived neurotrophic factor (GDNF); neurotrophin-3;neurotrophin-4; fibroblast growth factor (FGF) receptor; insulin-likegrowth factor (IGF); or a combination thereof.

XIII. Methods of Light Induced Nerve Ablation

The present disclosure provides methods of delivering a photosensitizingagent to a human neuron or nerve comprising: contacting the human neuronor nerve with a human neuron or nerve targeting molecule comprising (a)a peptide that specifically binds to the neuron or nerve, or componentof either, and (b) a photosensitizing agent. In some embodiments, themethod further comprises exposing the human neuron or nerve with a lightsource that activates the photosensitizing agent, wherein the activatedphotosensitizing agent induces ablation or killing of the human neuronor nerve. Upon exposure to a specific wavelength of light, aphotosensitizing agent reacts with molecular oxygen to produce singletoxygen, which is cytotoxic. In certain embodiments, a photosensitizingagent is a porphyrin, chlorin, or dye. Examples of photosensitizingagents include porphyrin, protoporfin IX, purlytin, verteporfin, HPPH,temoporfin, methylene blue, photofrin, protofrin, hematoporphyrin,Talaporfin, benzopophyrin derivative monoacid, 5-aminileuvolinic acid,Lutetium texaphyrin, metallophthalocyanine,metallo-naphthocyaninesulfobenzo-porphyrazines,metallo-naphthalocyanines, zinc tetrasulfophthalocyanine,bacteriochlorins, metallochlorins, chlorine derivative,Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein(DBF), IR700DX, naphthalocyanine, and porphyrin derivatives. In someembodiments, the human neuron or nerve targeting molecule comprises apeptide sequence comprising SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGClinker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22), EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS(HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), or5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the peptide comprises: SGQVPWEEPYYVVKKSS (HNP 401;SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), or DLPDIIWDFNWETA(HNP 403; SEQ ID NO:3). In some embodiments the peptide comprises:SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC(HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC(HNP401-N-2 with GGC linker; SEQ ID NO:7), SGQVPWEEPYYVVKK (HNP401-C-2;SEQ ID NO:25), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), orQVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21). In someembodiments the peptide comprises SGQVPWEEPYYVVKKSS (HNP 401; SEQ IDNO:1). In some embodiments the peptide comprises WEYHYVDLNWTSQHPQ (HNP402; SEQ ID NO:2). In some embodiments the peptide comprisesDLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments the peptidecomprises Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).In some embodiments the peptide comprises Ac-WEYHYVDLNWTSQHPQGGC (HNP402with GGC linker; SEQ ID NO:5). In some embodiments the peptide comprisesAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In someembodiments the peptide comprises QVPWEEPYYVVKKSS (HNP401-N-2; SEQ IDNO:20). In some embodiments the peptide comprises QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21). In some embodiments thepeptide comprises Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQID NO:7). In some embodiments the peptide comprises PWEEPYYVVKKSS(HNP401-N-4; SEQ ID NO:22). In some embodiments the peptide comprisesAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In someembodiments the peptide comprises EEPYYVVKKSS (HNP401-N-6; SEQ IDNO:23). In some embodiments the peptide comprises Ac-EEPYYVVKKSSGGC(HNP401-N-6 with GGC linker; SEQ ID NO:9). In some embodiments thepeptide comprises PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In someembodiments the peptide comprises Ac-PYYVVKKSSGGC (HNP401-N-8 with GGClinker; SEQ ID NO:10). In some embodiments the peptide comprisesSGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some embodiments thepeptide comprises Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQID NO:11). In some embodiments the peptide comprises SGQVPWEEPYYVV(HNP401-C-4; SEQ ID NO:26). In some embodiments the peptide comprisesAc-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12). In someembodiments the peptide comprises SGQVPWEEPYY (HNP401-C-6; SEQ IDNO:27). In some embodiments the peptide comprises Ac-SGQVPWEEPYYGGC(HNP401-C-6 with GGC linker; SEQ ID NO:13). In some embodiments thepeptide comprises SGQVPWEEP (HNP401-C-8; SEQ ID NO:28). In someembodiments the peptide comprises 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2with GG linker; SEQ ID NO:104). In some embodiments the peptidecomprises Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14). Insome embodiments the peptide comprises DTHAHAKPRVPAFKSV (HNP 404; SEQ IDNO:16). In some embodiments, the targeting molecule comprises a peptidethat is not Ac-SHSNTQTLAKAPEHTGC (Ac-NP41 with GC linker; SEQ ID NO:17).In some embodiments, the targeting molecule comprises a peptide that isnot SHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID NO:18). In someembodiments, the targeting molecule comprises a peptide that is notNTQTLAKAPEHT (SEQ ID NO:19).

Human neuron or nerve targeting molecules comprising a photosensitizingagent as disclosed herein can be used in methods of localized nervekilling in a subject. In some embodiments, human neuron or nervetargeting molecules comprising a photosensitizing agent are used fortreating chronic pain (e.g., back, neck, or joint pain) in subject. Insome embodiments, human neuron or nerve targeting molecules comprising aphotosensitizing agent are used for treating prostate cancer in asubject. Autonomic innervation may contribute to prostate cancer growthand metastasis by light induced ablation of local autonomic nerves. Thuslocal autonomic nerves may be a viable target for prostate cancertherapy. In some embodiments, human neuron or nerve targeting moleculescomprising a photosensitizing agent are used for treating renovascularhypertension in a subject by light induced ablation of sympatheticnerves in the renal vessels. In some embodiments, human neuron or nervetargeting molecules comprising a photosensitizing agent are used fortreating excessive sweating. In some embodiments, human neuron or nervetargeting molecules comprising a photosensitizing agent are used fortreating cardiac arrhythmias. In some embodiments, human neuron or nervetargeting molecules comprising a photosensitizing agent are used fortreating pathological muscle spasms (e.g., Meige syndrome, hemifacialspasm, torticollis).

XIV. Pharmaceutical Compositions

Disclosed herein, in certain embodiments, are pharmaceuticalcompositions comprising a human neuron or nerve targeting moleculedisclosed herein. Pharmaceutical compositions herein are formulatedusing one or more physiologically acceptable carriers includingexcipients and auxiliaries which facilitate processing of the activeagents into preparations which are used pharmaceutically. Properformulation is dependent upon the route of administration chosen. Asummary of pharmaceutical compositions is found, for example, inRemington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton,Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington'sPharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975;Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,Marcel Decker, New York, N. Y., 1980; and Pharmaceutical Dosage Formsand Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins,1999). In some embodiments, the human neuron or nerve targeting moleculecomprises a peptide sequence selected from the group consisting of:SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG(HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22), EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS(HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25),SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker;SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ IDNO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG(HNP401-C-8 with GG linker; SEQ ID NO:124), and5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).In some embodiments the targeting molecule comprises a peptide selectedfrom the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403;SEQ ID NO:3). In some embodiments, the targeting molecule comprises apeptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker;SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ IDNO:7), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS(HNP401-N-2; SEQ ID NO:20), and QVPWEEPYYVVKKSSGG (HNP401-N-2 with GGlinker; SEQ ID NO:21). In some embodiments, the targeting moleculecomprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In someembodiments, the targeting molecule comprises the peptideWEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some embodiments, thetargeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ IDNO:3). In some embodiments, the targeting molecule comprises the peptideAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In someembodiments, the targeting molecule comprises the peptideAc-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5). In someembodiments, the targeting molecule comprises the peptideAc-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In someembodiments, the targeting molecule comprises the peptideQVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, thetargeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2with GG linker; SEQ ID NO:21). In some embodiments, the targetingmolecule comprises the peptide Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 withGGC linker; SEQ ID NO:7). In some embodiments, the targeting moleculecomprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In someembodiments, the targeting molecule comprises the peptideAc-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In someembodiments, the targeting molecule comprises the peptide EEPYYVVKKSS(HNP401-N-6; SEQ ID NO:23). In some embodiments, the targeting moleculecomprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQID NO:9). In some embodiments, the targeting molecule comprises thepeptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In some embodiments, thetargeting molecule comprises the peptide Ac-PYYVVKKSSGGC (HNP401-N-8with GGC linker; SEQ ID NO:10). In some embodiments, the targetingmolecule comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ IDNO:25). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ IDNO:11). In some embodiments, the targeting molecule comprises thepeptide SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26). In some embodiments,the targeting molecule comprises the peptide Ac-SGQVPWEEPYYVVGGC(HNP401-C-4 with GGC linker; SEQ ID NO:12). In some embodiments, thetargeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ IDNO:27). In some embodiments, the targeting molecule comprises thepeptide Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). Insome embodiments, the targeting molecule comprises the peptide SGQVPWEEP(HNP401-C-8; SEQ ID NO:28). In some embodiments, the targeting moleculecomprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GGlinker; SEQ ID NO:104). In some embodiments, the targeting moleculecomprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQID NO:14). In some embodiments, the targeting molecule comprises thepeptide DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16). In some embodiments,the targeting molecule comprises a peptide that is notAc-SHSNTQTLAKAPEHTGC (Ac-NP41 with GC linker; SEQ ID NO:17). In someembodiments, the targeting molecule comprises a peptide that is notSHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID NO:18). In someembodiments, the targeting molecule comprises a peptide that is notNTQTLAKAPEHT (SEQ ID NO:19).

In certain embodiments, a pharmaceutical composition disclosed hereinfurther comprises a pharmaceutically acceptable diluent(s),excipient(s), or carrier(s). In some embodiments, the pharmaceuticalcompositions include other medicinal or pharmaceutical agents, carriers,adjuvants, such as preserving, stabilizing, wetting or emulsifyingagents, solution promoters, salts for regulating the osmotic pressure,and/or buffers. In addition, the pharmaceutical compositions alsocontain other therapeutically valuable substances.

In certain embodiments, the human neuron or nerve targeting moleculesdisclosed herein are delivered to a subject via a drug delivery vehicleor carrier. In some embodiments, a delivery vehicle is made from naturalor synthetic materials or both. In some embodiments, a delivery vehicleis a nanoparticle, microparticle, polymeric micelle, nanocapsule,dendrimer, large PEG, nanogel, liposome, fullerene, nanostructured lipidcarrier, nanoshell, quantum dot, protein-based nanocarriers (e.g.,albumin, elastin, gliadin, legumin, zein, soy protein, milk protein,whey based nanocarriers), organic nanocarrier (e.g., gelatin, dextran,guar gum, chitosan, collagen), polysaccharide based carrier (e.g.,dextran, chitosan, pectin), lipid emulsion, or a combination thereof.

In certain embodiments, a pharmaceutical composition disclosed herein isadministered to a subject by any suitable administration route,including but not limited to, parenteral (intravenous, subcutaneous,intraperitoneal, intramuscular, intravascular, intrathecal,intravitreal, infusion, or local) administration.

Formulations suitable for intramuscular, subcutaneous, or intravenousinjection include physiologically acceptable sterile aqueous ornon-aqueous solutions, dispersions, suspensions or emulsions, andsterile powders for reconstitution into sterile injectable solutions ordispersions. Examples of suitable aqueous and non-aqueous carriers,diluents, solvents, or vehicles including water, ethanol, polyols(propyleneglycol, polyethylene-glycol, glycerol, cremophor and thelike), suitable mixtures thereof, vegetable oils (such as olive oil) andinjectable organic esters such as ethyl oleate. Proper fluidity ismaintained, for example, by the use of a coating such as lecithin, bythe maintenance of the required particle size in the case ofdispersions, and by the use of surfactants. Formulations suitable forsubcutaneous injection also contain optional additives such aspreserving, wetting, emulsifying, and dispensing agents.

For intravenous injections, an active agent is optionally formulated inaqueous solutions, preferably in physiologically compatible buffers suchas Hank's solution, Ringer's solution, or physiological saline buffer.

Parenteral injections optionally involve bolus injection or continuousinfusion. Formulations for injection are optionally presented in unitdosage form, e.g., in ampoules or in multi dose containers, with anadded preservative. In some embodiments, the pharmaceutical compositiondescribed herein are in a form suitable for parenteral injection assterile suspensions, solutions or emulsions in oily or aqueous vehicles,and contain formulatory agents such as suspending, stabilizing and/ordispersing agents. Pharmaceutical formulations for parenteraladministration include aqueous solutions of an active agent in watersoluble form. Additionally, suspensions are optionally prepared asappropriate oily injection suspensions.

In some embodiments, the pharmaceutical composition described herein isin unit dosage forms suitable for single administration of precisedosages. In unit dosage form, the formulation is divided into unit dosescontaining appropriate quantities of an active agent disclosed herein.In some embodiments, the unit dosage is in the form of a packagecontaining discrete quantities of the formulation. Non-limiting examplesare packaged tablets or capsules, and powders in vials or ampoules. Insome embodiments, aqueous suspension compositions are packaged insingle-dose non-reclosable containers. Alternatively, multiple-dosereclosable containers are used, in which case it is typical to include apreservative in the composition. By way of example only, formulationsfor parenteral injection are presented in unit dosage form, whichinclude, but are not limited to ampoules, or in multi dose containers,with an added preservative.

In some embodiments, the human neuron or nerve targeting molecule isadministered via systemic intravenous injection into human patients.

EXAMPLES Example 1: Peptides for Targeting Human Nerves and their Use inImage Guided Surgery, Diagnostics and Therapeutic Delivery SUMMARY

Phage display screens to identify peptides that bind human nerves andcould therefore be useful for systemic in-vivo labeling of nerves duringfluorescence assisted surgery was used. Specifically, m13 phagelibraries expressing 16 random amino acid sequences on the N-terminus ofgill (Creative Biolabs) were processed through selections for binding tofreshly resected or frozen human nerves. In parallel, a newly designedNP41 X12+4 library was screened. Each library was processed through upto 6 binding and wash cycles. Selected phage were additionally selectedfor counter-selected for low affinity muscles and fat tissue bypreabsobing library, And any high affinity binder, with tissue prior topositive selection for nerve binding. Sequencing of individual phageyielded these unique sequences that were highly enriched and thereforehigher affinity relative to the pool of clones: SGQVPWEEPYYVVKKSS(HNP401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP402; SEQ ID NO:2),DLPDIIWDFNWETA (HNP403; SEQ ID NO:3) from the X16 library andDTHAHAKPRVPAFKSV (HNP404; SEQ ID NO:16) from NP41-X12+4 library. Aminoacid sequences derived from sequences of selected phage were chemicallysynthesized as peptides by solid-phase synthesis and labeled withfluorescein (FAM) or Cy5 at the C-terminus via a GGC linker for in-vitrobinding to human nerves and in-vivo labeling of rodent nerves. Stronglabeling of freshly sections of human nerves and in-vivo labeled mousesciatic nerves was shown. Useful labeling occurs between 2-6 hours afterintravenous administration and could be visualized using a customizedfluorescence dissecting microscope, a Maestro imager from CRI, or aZeiss Lumar.

Preservation of peripheral nerves is one of the most important goals ofany surgical procedure, because accidental transection of peripheralnerves during surgical procedures lead to significant morbidity forpatients. Also, nerves grow back more slowly and incompletely aftertransection than almost any other tissue. Typically, peripheral nervesare identified by their relatively constant relationship to nearbystructures as well as by their typical appearance of being elongatedwhitish, glistening structures. However, in many instances,identification of peripheral nerves using these criteria can bedifficult: for example in cases of tumor involvement, in instances ofinflammation/infection, in a previously operated surgical field, or whenthe nerve is encased in bone.

Current methods for nerve labeling primarily depend on retrograde oranterograde tracing of individually identified axonal tracts via the useof fluorescent dyes. The fluorescent dyes are either applied to theinnervation target and travel in a retrograde fashion to label theinnervating nerve fibers, or are applied directly to the identifiednerves and label the nerve fibers both anterogradely and retrogradely.This technique has the drawback of being able to label only one nervefiber tract at a time, depending on where the dye has been injected. Asecond drawback is the limited accumulation of fluorescent dyes alongthe axonal tracts, because retrograde axonal tracers typicallyaccumulate in the neural cell body and axonal labeling with thesefluorescent dyes is limited. A third disadvantage of this technique isthat retrograde transport is relatively slow (on the order ofmillimeters per day) and therefore takes a long time to label humannerves, which are often longer than a meter (as in the case of thesciatic nerve and its arborizations). Furthermore, the application offluorescent dyes to innervation targets such as direct intramuscularinjections to label motor nerves is typically messy with a variableamount of the tracer dye remaining at the injection site. As dissectionof nerves depends on accurate visualization of adjacent structures priorto encountering them, a surgical site that is contaminated withfluorescent dyes would not be desirable. Finally, the direct injectionof the fluorescent dye itself may be damaging to the target organs ornerve of interest, either by mechanical damage or by the very high localconcentration of dye and vehicle at the injection site.

The method of systemic injection of fluorescently labeled peptides tolabel nerves described in this document addresses all of thedisadvantages of fluorescently tracers described above. First, as thepeptides are delivered systemically, all peripheral nerves in the bodyhave the potential of being labeled. This is contrast to the labeling ofonly one nerve at a time as with current methods. Second, as thepeptides described here were selected for their ability to bind nerves,the nerve fibers are clearly visualized compared to adjacent non-neuralstructures. This is in contrast to the preferential accumulation toneural cell bodies rather than axonal processes with most currentfluorescent dyes. Third, the binding of the peptides described here tonerves occurs very quickly and visualization of peripheral nerves usingthis technique can be accomplished within hours. This is in contrast tothe relatively slow rate of labeling with anterograde or retrogradetracers. Finally, since the peptides are applied systemically viaintravenous injection, damage to nerves at the injection site is not anissue.

Nerve-homing peptide sequences that were derived using mouse peripheralnerves for laboratory research have been previously described (U.S. Pat.No. 8,685,372, Apr. 1, 2014). However, because the intended eventualclinical application of nerve labeling is in human patients,identification of unique peptide sequences that bind human nerves wassought. The peptide sequences described in this application wasidentified by their ability to bind human nerves. These peptidesequences were identified by their ability to bind human nerves,following systemic intravenous injection into human patients, and assuch these peptides will be much more likely to bind human nervescompared to sequences that were selected against rodent nerves.

Current methods for labeling nerves involve the application offluorescent tracer dyes (Fast Blue, Rhodamine-isothiocyanate,Fluoro-Ruby, Fluoro-Emerald), carbocyanine dyes (DiI, DiAsp, DiO, DiA),Fluoro-Gold, fluorescently labeled latex beads, fluorescently labeledplant lectins and bacterial toxins (wheat germ agglutinin, peanutagglutinin, concanavalin A, Phaseolus vulgaris-leucoagglutininin(PHA-L), soybean agglutinin, Ulex europaeus agglutinin, Ricinus communisagglutinin (I and II), tetanus toxin fragment C, cholera toxin B andfluorescently labeled dextran conjugates.

Methods: Experimental Details:

m13 phage libraries expressing random 16 amino acid sequences on theN-terminus of gill (Creative Biolabs) or an internally derived libraryexpressing derivative of NP41 were used to identify peptides that bondhuman nerve tissue.

Selection of Peptides

Human peripheral nerves were obtained from patients undergoing nerveresection procedures and homogenized. Phage library mixture wasincubated with nerve homogenate or nerve homogenates that had been boundto high protein binding 6 well plates. Following incubation, the mixturewere either centrifuged and the pellet washed with PBS, or plate waswashed with PBS. The pellet was rehomogenized and plated for titer andre-amplification or released from plate with low pH buffer. Phage thatwere bound at each round were sequenced and repeats noted. No repeatswere identified until round 4 of selection.

SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP402; SEQID NO:2), DLPDIIWDFNWETA (HNP403; SEQ ID NO:3) from the X16 library andDTHAHAKPRVPAFKSV (SEQ ID NO:16) were identified after round 5, Table 1

TABLE 1 Peptides Identified Number SEQ Repeating of ID Name sequencesrepeats NO: HNP 401 SGQVPWEEPYYVVKKSS >10  1 HNP 402WEYHYVDLNWTSQHPQ >10  2 HNP 403 DLPDIIWDFNWETA   7  3 HNP 404DTHAHAKPRVPAFKSV   3 16

In Vivo Testing of Peptides

Either 150 or 450 nmoles nmoles of fluorescein labeled syntheticpeptides were injected intravenously into mice. After a 2 hour waitingperiod for washout of nonspecific binding, mice were anesthetized andskin incisions were made over the dorsal surface of the hind legs toexpose the sciatic nerve. Brightfield and fluorescent images wereobtained with a dissecting microscope using Metamorph software (FIG. 1).Quantitation of fluorescence of nerves and adjacent non-nerve tissue wasperformed with Image J (FIG. 2). Peptides were also topically applied tohuman nerve sections. Nerves were freshly frozen in OTC prior tosectioning. Peptides were topically applied at concentration at 300 uMwith images being shown for HNP401, HNP402, HNP404 and previous reportednerve binding peptides NP41 (FIG. 3)). Also shown are images forvariants HNP301 (SEFPRSWDMETN) and NP124. NP713 was also tested and hasnot not reported in a publication. NP713, is a derivative of NP41 withsequence NTHPHTTSRVPSQIAR that was enriched after 7 rounds of selectionagainst mouse tissue, and was also found after 4 rounds of selectionagainst human tissue. Binding of NP713 phage compared to wildtype phageshowed a 4.8-fold higher nerve:muscle ratio. FAM-NP713 showed similarnerve:muscle contrast to NP41 (Data not shown here). All D-amino acidcontrols for NP-41 and NP713 and non peptide conjugatedcarboxyfluorescein, are also shown. HNP401 shows the highest nervespecific contrast with the majority of the labelling occurring in theperineurium. To further demonstrate HNP401 selective binding, HNP401,NP41 and HNP404 were tested at a lower concentration, 100 μM (FIG. 4).HNP401 was then tested for labeling of rat sciatic and rat prostatecavernosal nerve in-vivo. FIG. 5 shows in-vivo labeling of rat sciaticnerve. FIGS. 6 and 7 show in-vivo labeling of rat prostate cavernosalnerve with comparison to white light visualization.

Nerve-homing peptides sequences that were identified by their ability tobind mouse nerves for laboratory research were previously described.Because the peptide sequences described in this document were identifiedby their ability to bind human nerves, following systemic intravenousinjection into human patients, these peptides will be much more likelyto bind human nerves compared to sequences that were selected againstrodent nerves.

Fluorescently labeled human nerve-binding peptides are appliedsystemically via intravenous injection. Following a short waiting periodfor washout of nonspecific binding, peripheral nerves can be visualizedwithin a surgical field with appropriate excitation and emissionfilters.

Human nerve-binding peptides might also be conjugated to factors thatmay have neurotrophic or protective properties to nerves. Followingsystemic application via intravenous injection,peptide-trophic/neuroprotective factor conjugates might facilitaterepair/regeneration of damaged nerves both in the periphery and in thespinal cord.

Human nerve binding peptides conjugated to neuroprotective/neurotrophicfactors may also be conjugated to injury homing peptides to furtherimprove localized delivery of these factors to injured nerves,potentially faciliting resistance to injury/repair/regeneration.

Applications & Uses:

Fluorescently labeled nerve-binding peptides can be used to assistsurgeons in the visualization of nerves during surgical procedures priorto physically encountering and thus potentially damaging them. This isparticularly important during surgery on the prostate gland, because thecavernosal nerves controlling male erections run very near the prostatebut are practically invisible ordinarily.

Nerve binding peptide-neurotrophic/neuroprotective factor conjugates canbe used to facilitate repair/regeneration of damaged nerves.

Nerve binding peptides could be conjugated to photosensitizing dyes forpotential use with light induced nerve killing as a treatment forlocalized pain

REFERENCES

-   1. Whitney M, Crisp J, Nguyen L, Friedman B, Gross L, Steinbach P,    Tsien R, Nguyen Q. Fluorescentpeptides highlight peripheral nerves    during surgery in mice. Nature Biotechnology. 2011; 29:352-356-   2. Wu A P, Whitney M A, Crisp J L, Friedman B, Tsien R Y, Nguyen    Q T. Improved facial nerve identification with novel fluorescently    labeled probe. The Laryngoscope. 2011; 121:805-810-   3. Kobbert C., Apps, R., Bechmann, I., Laciego, J. L., Mey, J.,    Thanos, S. Currents concepts in neuroanatomical tracing. Progress in    Neurobiology 62 (2000) 327-351.-   4. Richmond, F. J. R., Gladdy R., Creasy, J. L., Ktamura S., Smits,    E., Thomson D. B. Efficacy of seven retrograde tracers, compared in    multiple-labelling studies of feline motoneurones. Journal of    Neuroscience Methods 53 (1994) 35-46.2-   5. Marangos, N., Illing R., Kruger J., Laszig R. In vivo    visualization of the cochlear nerve and nuclei with fluorescent    axonal tracers. Hearing Research 162 (2001) 48-52.-   6. O'Malley, M, Wittkopf, J., Cutler J., Labadie, R, Hackett, T,    Haynes, D. Fluorescent retrograde axonal tracing of the facial    nerve. The Laryngoscope 116 (2006) 1792-1797.

Example 2: Optimized Peptides for Targeting Human Nerves and their Usein Image Guided Surgery, Diagnostics and Therapeutic Delivery SUMMARY

Phage display screens were used to identify peptides that bind humannerves and could therefore be useful for systemic in-vivo labeling ofnerves during fluorescence assisted surgery. Specifically, m13 phagelibraries expressing 16 random amino acid sequences on the N-terminus ofgill (Creative Biolabs) were processed through selections for binding tofreshly resected or frozen human nerves. Library was processed throughup to 6 binding and wash cycles. Selected phage were additionallyselected for counter-selected for low affinity muscles and fat tissue bypreabsobing library, And any high affinity binder, with tissue prior topositive selection for nerve binding. Sequencing of individual phageyielded these unique sequences that were highly enriched and thereforehigher affinity relative to the pool of clones: SGQVPWEEPYYVVKKSS(HNP401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP402; SEQ ID NO:2),DLPDIIWDFNWETA (HNP403; SEQ ID NO:3). Amino acid sequences derived fromsequences of selected phage were chemically synthesized as peptides bysolid-phase synthesis and labeled with fluorescein or Cy5 for in-vitrobinding to human nerves and in-vivo labeling of rodent nerves. Stronglabeling of freshly sections of human peripheral nerves (motor, sensory,autonomic) and in-vivo labeled mouse and rat sciatic nerves was shown.Useful labeling occurs between 2-6 hours after intravenousadministration and could be visualized using a customized fluorescencedissecting microscope, a Maestro imager from CRI, or a Zeiss Lumardissecting microscope.

Results Nerve Identification and Preservation is Essential in Surgery ofthe Head and Neck.

Identification of peripheral nerves is critical for their preservationduring surgery, because accidental transection or injury can lead tosignificant patient morbidity including chronic pain, numbness orpermanent paralysis¹. Nerve identification is especially importantduring surgery of the head and neck. For example, facial nervedysfunction has been reported to be as high as 40% during the acutepostoperative period and 30% at 1 month following parotidectomy^(2,3).Similarly, facial nerve dysfunction has been reported to be as high as30% at 1 year following vestibular schwannoma surgery⁴. Temporary andpermanent vocal fold immobility are major surgical complications ofthyroid surgery, anterior cervical approaches to the spine,esophagectomy and carotid endarterectomy⁵. Although the course of thefacial nerve typically follow defined anatomical landmarks, extensivepatient to patient variability has been documented for every branch ofthe extratemporal facial nerve including variability in total number ofdivisions, origin of individual divisions and connections betweendivisions⁶⁻⁹. Even within the same patient, the left and right facialnerve may display differences in course and divisions¹⁰. Similarvariability has been documented for the recurrent laryngealnerve^(11,12). In instances of tumor invasion, inflammation, trauma orrepeat surgery, nerve identification can be even more challenging.Finally, identification of degenerated nerves, which are criticallyimportant during reconstructive surgery, is even more difficult thantheir functioning counterpart as they become smaller and thinner overtime. Consequently, any means of improving the visual determinationbetween nerve and non-nerve tissue would represent a significant advancein surgical technique.

Nerve Identification and Preservation is Essential During OtherSurgeries Including Prostate Cancer Surgery.

Prostate cancer is the most common solid organ malignancy in U.S. men.For men with localized prostate cancer surgery results in excellentcancer control. All too often this cancer treatment comes at the expenseof erectile function, urinary control, and overall quality of life.Preservation of the autonomic neurovascular bundles during radicalprostatectomy is an important aspect of the operation. For nearly 20years the importance of preserving the autonomic nerves that run alongthe posterolateral aspect of the prostate in order to preserve erectilefunction has been recognized. The autonomic nerve fibers themselves arerarely visualized, however. Instead surgeons preserve the blood vesselcomplexes, or neurovascular bundles, that have been shown to have thehighest density of autonomic nerves. The exact position and distributionof these autonomic nerves are variable¹³⁻¹⁸ as even in the mostexperienced hands, erectile dysfunction and urinary incontinence arecommon¹⁴. Improved sexual function outcomes are associated withincreasing surgeon experience and avoidance of crush or tractioninjuries on these nerves. Significant anatomic variation and differencesin surgeon experience and volume create an opportunity to improvesurgical quality while minimizing adverse outcomes. In instances oftumor invasion, inflammation, trauma or repeat surgery, nerveidentification and preservation would represent an additional challenge.Finally, the growing use of robotically-assisted surgery, with itsinherent lack of haptic feedback¹⁹, further increases the surgeon'sdependence on visual information. Consequently, any means of improvingthe visual determination between nerve and non-nerve tissue wouldrepresent a significant advance.

Small Nerves are Hard to Identify During Surgery.

Thin or buried nerves are particularly difficult to distinguish and aretherefore the most likely to be damaged during surgical procedures.Identification of motor nerves prior to direct exposure is currentlydependent on electromyographic (EMG) monitoring²⁰⁻²² in which astimulating electrode is inserted and distal muscle twitches aremonitored. EMG is not an imaging technique, so even if a nerve has beenidentified in one location there is no visual guidance as to how farfrom the stimulation site and in which direction the nerve lies.Furthermore, EMG only identifies motor pathways, not sensory fibers suchas the first two divisions of the trigeminal nerve or thecochleovestibular nerve, nor sympathetic tracts such as theneurovascular bundle surrounding the prostate gland²³⁻²⁵, where nerveinjury following radical prostatectomy leads to significant urinaryincontinence and erectile dysfunction²⁶. Electrode insertion may itselfdamage a nerve. Finally, EMG fails if axonal or neuromusculartransmission is temporarily blocked distal to the recording site bynerve compression, trauma, tumor invasion, local anesthetics, orneuromuscular blockers. There are some potential technologies for invivo nerve visualization without exogenous probe molecules, such asoptical coherence tomography²⁷ or laser confocal microscopy²⁸. However,nerves have very little intrinsic contrast to distinguish them fromother tissues, and these techniques do not readily produce real-timelive images over the field of view necessary for guiding surgery.Degenerated nerves, important to identify during reconstructive surgeryafter cancer resection, traumatic or therapeutic amputations, would alsohave no myelin and therefore would not benefit from these agents.

Competing Strategies to Improve Nerve Visualization During Surgery.

For these reasons, there is much interest in development of labelingreagents to improve nerve visualization during surgery. There has beenfocus on nerve labeling which depend on retrograde or anterogradetracing of individually axonal tracts via the use of fluorescentdyes²⁹⁻³² or the B subunit of Cholera toxin (CTb488)³³. The fluorescentdyes are either applied to the innervation target and travel in aretrograde fashion to label the innervating nerve fibers, or are applieddirectly to the identified nerves and label the nerve fibers bothanterogradely and retrogradely. Local injections have the drawback ofbeing able to label only one nerve fiber tract at a time. Anterogradeand retrograde transport is relatively slow and can take days to travela few millimeters, while leaving most of the tracer at the injectionsite. As dissection of nerves depends on accurate visualization ofadjacent structures, a surgical site that is heavily contaminated withexcess fluorescent dyes would not be desirable. Finally, the directinjection of the fluorescent dye may be damaging to the target organs ornerve of interest, either by mechanical damage or by the very high localconcentration of dye and vehicle at the injection site.

More recently, there has interest in using vascular dyes such asindocyanine green (ICG) to label the vascular supply of nerves (i.e.vaso nervorum)^(34,35). One limitation of this technology is that smallnerves (such as cavernosal nerves important for prostate surgery) willhave proportionally less vaso nervorum, limiting contrast and intensitycompared to adjacent tissue.

There has also been focus on agents targeting myelin includingdistyrylbenzene (DSB) derivatives³⁶, coumarin derivatives andanti-ganglioside antibodies³⁷. DSB and coumarin derivatives are smallmolecules with intrinsic fluorescence while anti-ganglioside antibodiesare conjugated to fluorescent dyes for imagine³⁶⁻⁴³. While thesemolecules are potentially promising for peripheral nerve imaging,non-myelinated nerves such as cavernosal nerves (which are autonomic andminimally myelinated) would likely have little binding, thereby limitingtheir utility in these important surgeries. Degenerated nerves wouldalso have limited myelin present and thus would not be labeled withthese agents.

The method of systemic injection of fluorescently labeled peptides tolabel nerves described in this document addresses all of thedisadvantages of other nerve targeting techniques described above.First, as the peptides are delivered systemically, all peripheral nervesin the body have the potential of being labeled. This is contrast to thelabeling of only one nerve at a time as with current methods. Second, asthe peptides described here were selected for their ability to bindnerves, the nerve fibers are clearly visualized compared to adjacentnon-neural structures. This is in contrast to the preferentialaccumulation to neural cell bodies rather than axonal processes withmost current fluorescent dyes. Third, the binding of the peptidesdescribed here to nerves occurs very quickly and visualization ofperipheral nerves using this technique can be accomplished within hours.This is in contrast to the relatively slow rate of labeling withanterograde or retrograde tracers. Finally, since the peptides areapplied systemically via intravenous injection, damage to nerves at theinjection site is not an issue.

Nerve-homing peptide sequences that were derived using mouse peripheralnerves for laboratory research have been previously described (U.S. Pat.No. 8,685,372, Apr. 1, 2014 Peptides and aptamers for targeting ofneuron or nerves US20120148499 and WO2010121023A2). However, because theintended eventual clinical application of nerve labeling is in humanpatients, identification of unique peptide sequences that bind humannerves was sought. The peptide sequences described in this applicationwas identified by their ability to bind human nerves. These peptidesequences were identified by their ability to bind human nerves,following systemic intravenous injection into human patients, and assuch will be much more likely to bind human nerves compared to sequencesthan peptides that were selected against rodent nerves.

Current methods for labeling nerves involve the application offluorescent tracer dyes (Fast Blue, Rhodamine-isothiocyanate,Fluoro-Ruby, Fluoro-Emerald), carbocyanine dyes (DiI, DiAsp, DiO, DiA),Fluoro-Gold, fluorescently labeled latex beads, fluorescently labeledplant lectins and bacterial toxins (wheat germ agglutinin, peanutagglutinin, concanavalin A, Phaseolus vulgaris-leucoagglutininin(PHA-L), soybean agglutinin, Ulex europaeus agglutinin, Ricinus communisagglutinin (I and II), tetanus toxin fragment C, cholera toxin B andfluorescently labeled dextran conjugates.

The fluorescent dyes are either applied to the innervation target andtravel in a retrograde fashion to label the innervating nerve fibers, orare applied directly to the identified nerves and label the nerve fibersboth anterogradely and retrogradely. As mentioned above, localinjections have the drawback of being able to label only one nerve fibertract at a time. Anterograde and retrograde transport is relatively slowand can take days to travel a few millimeters, while leaving most of thetracer at the injection site. As dissection of nerves depends onaccurate visualization of adjacent structures, a surgical site that isheavily contaminated with excess fluorescent dyes would not bedesirable. Finally, the direct injection of the fluorescent dye may bedamaging to the target organs or nerve of interest, either by mechanicaldamage or by the very high local concentration of dye and vehicle at theinjection site.

More recently, there has interest in using vascular dyes such asindocyanine green (ICG) to label the vascular supply of nerves (i.e.vaso nervorum)^(34,35). One limitation of this technology is that smallnerves (such as cavernosal nerves important for prostate surgery) willhave proportionally less vaso nervorum, limiting contrast and intensitycompared to adjacent tissue.

There has also been focus on agents targeting myelin includingdistyrylbenzene (DSB) derivatives³⁶, coumarin derivatives andanti-ganglioside antibodies³⁷. DSB and coumarin derivatives are smallmolecules with intrinsic fluorescence while anti-ganglioside antibodiesare conjugated to fluorescent dyes for imagine³⁶⁻⁴³. While thesemolecules are potentially promising for peripheral nerve imaging,non-myelinated nerves such as cavernosal nerves (which are autonomic andminimally myelinated) would likely have little binding, thereby limitingtheir utility in these important surgeries. Degenerated nerves wouldalso have limited myelin present and thus would not be labeled withthese agents.

Nerve-homing peptides sequences that were identified by their ability tobind mouse nerves for laboratory research were previously described.Because the peptide sequences described in this document were identifiedby their ability to bind human nerves, following systemic intravenousinjection into human patients, these peptides will be much more likelyto bind human nerves compared to sequences that were selected againstrodent nerves.

Fluorescently labeled human nerve-binding peptides are appliedsystemically via intravenous injection. Following a short waiting periodfor washout of nonspecific binding, peripheral nerves can be visualizedwithin a surgical field with appropriate excitation and emissionfilters.

Human nerve-binding peptides might also be conjugated to factors thatmay have neurotrophic or protective properties to nerves. Followingsystemic application via intravenous injection,peptide-trophic/neuroprotective factor conjugates might facilitaterepair/regeneration of damaged nerves both in the periphery and in thespinal cord.

Human nerve binding peptides conjugated to neuroprotective/neurotrophicfactors may also be conjugated to injury homing peptides to furtherimprove localized delivery of these factors to injured nerves,potentially faciliting resistance to injury/repair/regeneration.

Methods Experimental Details:

m13 phage libraries expressing random 16 amino acid sequences on theN-terminus of gill (Creative Biolabs) or an internally derived libraryexpressing derivative of NP41 to identify peptides that bond human nervetissue were used. NTQTLAKAPEHT (NP-41; SEQ ID NO:15; see U.S. Pat. No.8,685,372 or International Patent Publication No. WO2010121023A2; bothof which are incorporated by reference herein in there entireties).

Selection of Peptides:

Human peripheral nerves were obtained from patients undergoing nerveresection procedures and homogenized. The phage library mixture wasincubated with nerve homogenate or nerve homogenates that had been boundto high protein binding 6 well plates. Following incubation, themixtures were either centrifuged and the pellet washed with PBS, orplate was washed with PBS. The pellet was rehomogenized and plated fortiter and re-amplification or released from plate with low pH buffer.Phage that were bound at each round were sequenced and repeats noted. Norepeats were identified until round 4 of selection.

The following peptides were identified: SGQVPWEEPYYVVKKSS (HNP401; SEQID NO:1), WEYHYVDLNWTSQHPQ (HNP402; SEQ ID NO:2), DLPDIIWDFNWETA(HNP403; SEQ ID NO:3) from the X16 library and DTHAHAKPRVPAFKSV (SEQ IDNO:16) were identified after round 5. See peptides Table 2.

TABLE 2 Peptides Identified Number SEQ Repeating of ID Name sequencesrepeats NO: HNP 401 SGQVPWEEPYYVVKKSS >10 1 HNP 402 WEYHYVDLNWTSQHPQ >102 HNP 403 DLPDIIWDFNWETA   7 3Demonstration of peptide binding to human nerves:

To determine the affinity of phage selected peptides for binding tohuman nerve they were chemically synthesized by solid-phase synthesisand labeled with fluorescein at the C-terminus via a GGC linker.Peptides were topically applied to sectioned human sural nerve and humantemporalis muscle to determine nerve to muscle contrast. HNP401 showedthe highest binding and highlighting of human never (FIG. 8). Data forother peptides screened on human nerves and controls including freecarboxy fluorescein and NP41 screened on human nerve are additionallyshown. To confirm binding and contrast of HNP401 for additional nervetypes binding was compared in nerve and muscle with both HNP401 and NP41on facial brachial plexus nerve (FIG. 9). To quantify differentialbinding to human nerve versus muscle, fluorescence signal intensity wasmeasured for ROIs from the perineurium of select nerves and humantemporalis muscle that had been identically treated with topicalapplication of fluorescein labelled nerve binding peptides. FAM-HNP401showed selective binding to human sural nerve with 10.9× fluorescentsignal intensity (1374.44±425.96) compared to FAM-NP41 (126.17±61.03)(FIG. 9G), p=0.009, Student's t-test, unpaired). Nerve to musclecontrast was comparable at 3.03±0.57 for FAM-HNP401 and 2.28±0.96 forFAM-NP41 (FIG. 9H), p=0.236, Student's t-test, unpaired). Dose dependenttesting shows HNP401 has significant nerve binding down to 10 uM (FIG.10A-E) with increased nerve human binding of HNP401 compared to NP41detected at concentrations as high 375 uM (FIG. 10 F-I). FAM-HNP401 wasadditionally tested topically on ex-vivo tissue for labeling of mousefacial nerve with surrounding muscle (FIG. 10, J-M). Confocal imagingalso showed that HNP401-FAM binds epineurium, perineurium and endoneriumbut not axons (FIG. 10N).

In-vivo fluorescence imaging of sciatic nerve in mice that were injectedwith 450 nmols of FAM-HNP401 (FIG. 11A) or FAM-NP41 (FIG. 11B) showednerve contrast with HNP401 having 2.3 fold fluorescent intensitycompared to (NP41) (FIG. 11C) but with similar nerve to non-nervecontrast 5.79±0.81 for FAM-HNP401 and 6.63±1.63 for FAM-NP41 (FIG. 11D).FAM-HNP401 also highlighted rat prostate nerve (FIGS. 11 E-F) and ratsciatic nerve (FIG. 11G) at a dose of 2 μmole when imaged 3 hours postinjection or alternatively using a lower dose of 0.5 μmole HNP401 withimaging 10 mins after probe injection. Blood clearance of FAM-HNP401after injection of 100 nmols i.v. showed a half-life of 30 minutes whichis similar to FAM-NP41 (FIG. 11H).

FAM-HNP401 and FAM-NP41 were then tested topically for binding toautonomic nerves, isolated from excised prostate glands of two humanpatients (FIGS. 12 and 13). FAM-HNP401 (FIG. 12A) showed a significantlyhigher fluorescent signal in human autonomic nerves compared to FAM-NP41(FIG. 12B) at the same concentration. Labelled fiber were confirmed asnerve using anti-neurofilament antibody SMI312 (red) and DAPI (blue) toshow nuclear labeling (FIG. 12C). H&E staining was also done to confirmlabel tissue as nerve (FIG. 12D). Prostate nerve binding of HNP401 usingtissue from an additional patient is shown in FIG. 13. Similar stainingwas obtained for peripheral sensory anti-brachial cutaneous nerveisolated from a human arm (FIG. 12E-H).

Synthesis and Nerve Binding of Deletion Variants of HNP401

To optimize the HNP401 sequence, systematic deletion of 2 amino acidsfrom the C or N terminus (Table 3) followed by binding analysis to humannerve sections was performed. Deletion of amino acids from C terminalreduces binding efficacy and solubility. Removal of 2 amino acids fromthe N-termini improves the nerve binding with an average signalintensity of 1498.73 (+/−517.63) for N-2 and 744.63 (+/−130.18) forHNP401 [Student's t test, unpaired, 1 tail, p=0.07] (FIGS. 14 and 15).

TABLE 3 List of nerve binding peptides SEQ Unique ID PeptidePeptide sequence NO: HNP401 Ac-SGQVPWEEPYYVVKKSSGGC  4 (with GGC linker) HNP402 Ac-WEYHYVDLNWTSQHPQGGC  5 (with GGC  linker) HNP403Ac-DLPDIIWDFNWETAGGC  6 (with GGC  linker) HNP401-N-2Ac-QVPWEEPYYVVKKSSGGC  7 (with GGC  linker) HNP401-N-4Ac-PWEEPYYVVKKSSGGC  8 (with GGC  linker) HNP401-N-6 Ac-EEPYYVVKKSSGGC 9 (with GGC  linker) HNP401-N-8 Ac-PYYVVKKSSGGC 10 (with GGC  linker)HNP401-C-2 Ac-SGQVPWEEPYYVVKKGGC 11 (with GGC  linker) HNP401-C-4Ac-SGQVPWEEPYYVVGGC 12 (with GGC  linker) HNP401-C-6 Ac-SGQVPWEEPYYGGC13 (with GGC  linker) HNP401-C-8 Ac-SGQVPWEEPGGC 14 (with GGC  linker)

Applications & Uses:

Fluorescently labeled human nerve-binding peptides can be used to assistsurgeons in the visualization of nerves during surgical procedures priorto physically encountering and thus potentially damaging them. This isparticularly important when nerves are small, degenerated, invaded bycancer, injured by trauma or infection. For example, during surgery onthe prostate gland, the cavernosal nerves controlling male erections runvery near the prostate gland but are not definitively identified usingconventional light (white light reflectance) available in operatingtheaters.

Human nerve binding peptide-neurotrophic/neuroprotective factorconjugates can be used to facilitate repair/regeneration of damagednerves.

Human nerve binding peptides could be conjugated to photosensitizingdyes for potential use with light induced nerve killing as a treatmentfor chronic pain.

Human nerve binding peptides could be conjugated to photosensitizingdyes for potential use with light induced nerve killing as a treatmentfor excessive sweating.

Human nerve binding peptides could be conjugated to photosensitizingdyes for potential use with light induced nerve killing as a treatmentfor renovascular hypertension.

Human nerve binding peptides could be conjugated to photosensitizingdyes for potential use with light induced nerve killing as a treatmentfor cardiac arrhythmias.

Human nerve binding peptides could be conjugated to photosensitizingdyes for potential use with light induced nerve killing as a treatmentfor pathologic muscle spasms (Meige syndrome, hemifacial spasm,torticollis).

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Fluorescently-tagged anti-ganglioside antibody selectively    identifies peripheral nerve in living animals. Sci Rep. 2015;    5:15766.-   38. Gibbs S L, Xie Y, Goodwill H L, Nasr K A, Ashitate Y, Madigan V    J, Siclovan T_(M), Zavodszky M, Tan Hehir C A and Frangioni J V.    Structure-activity relationship of nerve-highlighting fluorophores.    PLoS One. 2013; 8:e73493.-   39. Cotero V E, Kimm S Y, Siclovan T_(M), Zhang R, Kim E M,    Matsumoto K, Gondo T, Scardino P T, Yazdanfar S, Laudone V P and Tan    Hehir C A. Improved Intraoperative Visualization of Nerves through a    Myelin-Binding Fluorophore and Dual-Mode Laparoscopic Imaging. PLoS    One. 2015; 10:e0130276.-   40. Cotero V E, Siclovan T, Zhang R, Carter R L, Bajaj A, LaPlante N    E, Kim E, Gray D, Staudinger V P, Yazdanfar S and Tan Hehir C A.    Intraoperative fluorescence imaging of peripheral and central nerves    through a myelin-selective contrast agent. Mol Imaging Biol. 2012;    14:708-17.-   41. 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Example 3: Nerve-Targeted Probes for Fluorescence-Guided IntraoperativeImaging Abstract

A fundamental goal of many surgeries is nerve preservation, asinadvertent injury can lead to patient morbidity including numbness,pain, localized paralysis and incontinence. Nerve identification duringsurgery currently relies on multiple parameters including anatomy,texture, color and relationship to surrounding structures to distinguishnerves from non-nerve tissues. Using white light illumination, which isthe standard in current operating rooms, the visual difference betweennerves and adjacent tissue can be imperceptible. A nerve-targeted probe,FAM-NP41, that binds to and highlights rodent motor and sensory nervesfollowing systemic administration was previously developed. Here it isdemonstrated that FAM-NP41 can highlight autonomic nerves within theprostate gland in living mice and rats with significant nerve tonon-nerve contrast in nerves as small as 50 μm in diameter.

To translate this methodology for potential clinical use in humanpatients, phage display was used to identify a novel peptide (HNP401)that selectively binds to human nerves. FAM-HNP401 can bind andhighlight multiple human peripheral nerves including lower leg suralnerve, upper arm medial antebrachial nerve and autonomic nervesincluding cavernosal nerve surrounding human prostate gland. The bindingdomain of HNP401 was identified by sequential deletion of amino acidsfrom the full-length peptide. HNP401 or an optimized variant could betranslated for use in a clinical setting for intraoperativeidentification of human nerves to improve visualization and potentiallydecrease the incidence of intra-surgical nerve injury.

Introduction

A fundamental goal of surgery is preservation of nerve function tominimize patient morbidity. Current nerve identification during surgeryutilizes non-quantifiable criteria such as anatomy, texture, color andrelationship to surrounding structures. In instances of trauma, tumorinvasion or infection, nerve identification using the above criteria canbe even more challenging. Using white light reflectance, the visualdifference between nerves, especially small nerves like the autonomicnerves within the prostate, and adjacent tissue can be imperceptible.Inadvertent injury to these thin or buried nerves is one of the mostmorbid but unintended consequence of surgery which can lead to loss offunction, numbness, and surgery induced neuropathic pain [1]. Forexample, radical prostatectomy (RP) can be performed for localizedprostate cancer with excellent locoregional control [2,3] However, evenwith nerve preserving radical prostatectomy there is a significant riskof erectile dysfunction and/or urinary incontinence, due to inadvertentinjury to autonomic nerves or the autonomic neurovascular bundles [4,5].Preservation of the autonomic neurovascular bundles along theposterolateral aspect of the prostate is an important aspect forfunctional preservation during RP. The autonomic nerve fibers themselvesare rarely visualized, but rather their position is presumed to trackalong vascular structures. The exact position and distribution of theseautonomic nerves are variable from patient to patient complicating theuse of anatomical location as the sole method of avoidance [6-8] andinjury can occur even in the most experienced hands. Recent studiesshowed that only 7% of RP patients regained pre-surgical state of fullerectile function in the first year [9] and 16% regained baselineerectile function 2 years after prostatectomy [10].

Tools to improve visualization of the neural structures in the prostatehave great potential for reducing morbidity from the radicalprostatectomy, as well as applications in many other nerve preservingsurgeries including cancer resection, trauma and reconstructiveprocedures. Systemic administration of a nerve imaging agent could allowthe labeling of all relevant nerves with a single probe administration.Previously, reported methods rely on retrograde or anterograde tracingof individually axonal tracts by direct application of fluorescent dyesto the innervation site [11, 12]. Styryl pyridinium dyes [13-15],aminostyryl dyes [16-18], oxazine 4 [19, 20], and anti-gangliosideantibodies [21] have been investigated in various preclinical models todetect motor, sensory and autonomic nerves.

A peptide sequence, Nerve Peptide 41 (NP41), was previously identifiedthrough phage display that preferentially binds and highlightsperipheral nerve tissue, enhancing visualization of motor and sensorynerves in live mice after systemic injection [22-24]. This peptide hasrelatively low affinity for nerve and rapid blood clearance (compared toantibodies) so it can be visualized a few hours after systemic injectionwith almost completely wash out by 24 hours [22]. NP41 has also beenshown to highlight degenerate nerves through the binding to structurallaminins in nerve fibers [24, 25]. We have now used this peptide forintraoperative identification of autonomic nerves in the prostate ofboth mice and rats. To allow clinical translation of nerve visualizationmethods for use in surgeries involving human patients, we have now usedphage display to identify a novel peptide HNP401 that, when labeled withfluorophore, selectively binds and highlights human nerves.Fluorescently labeled HNP401 can bind to and highlight human sensory andmotor nerves such as sural, medial antebrachial cutaneous, laryngeal,ansa cervicalis, great auricular nerve and autonomic nerves like thosewithin and around the prostate gland.

Results

To visualize the autonomic nerves within the prostate of mice, NP41peptide conjugated to fluorescein (FAM-NP41) was injected intravenouslyfollowed by imaging of prostate and surrounding tissue after surgicalresection. Strong fluorescence from dye that rapidly accumulates in thebladder hindered visualization of the nerves within the prostate. Toenhance visualization the bladder was surgically drained of urine andsutured prior to imaging. The urethra, an anatomically distinctstructure, is never emptied of urine as the mice are alive for theduration of the experiment, resulting in a continuous passage of urinecarrying metabolized peptide-dye to the bladder via the urethra. To aidfuture research, we have demonstrated using a fluorescent quenching dye(both directly injected in the bladder and through oral administration)to reduce high bladder fluorescence, as an alternative to surgicaldraining of the bladder (FIG. 21). FAM-NP41 was injected at dosesranging from 150-600 nmoles (^(˜)16 to 66 mg/kg) with a 600 nmoles(^(˜)30 nmol/g) dose showing optimal autonomic nerve contrast (FIGS.16A-16G). Low magnification fluorescent image show highlighting of asingle nerve fiber running adjacent to the urethra (FIG. 16A). The nerveis extremely faint in a high magnification image using white lightreflectance (FIG. 16B) but becomes distinctly visible with FAM-NP41labeling (FIG. 16C). To quantify nerve detection a total of 10 mice wereinjected with 600 nmoles FAM-NP41 and signal intensity was measured fornerve versus adjacent non-nerve tissue using both fluorescence and whitelight reflectance. Values to the right of the line indicate that thereis improved visualization with fluorescence compared to reflected light.Average nerve to non-nerve signal intensity with fluorescent guidancewas 1.256±0.14 (n=12, p<0.001) compared to 1.086±0.07 (n=12) forwhite-light reflectance (FIG. 16D).

Because prostate nerves in mice were very small and challenging to image(i.e. requiring high dose of FAM-NP41) we extended our study to thevisualization of autonomic nerve within the prostate of rats. Tovisualize autonomic nerves in male Sprague Dawley rats, FAM-NP41 wasinjected intravenously at a dose of 12 nmol/gram, followed by imaging.This is a 2.5× lower dose relative to weight compared to the 600 nmolsused in 20 gram mice. Useful labeling occurred 2 to 6 hours afterintravenous administration which was visualized using a customizedfluorescence dissecting microscope. FAM-NP41 nerve highlighting enablesvisualization of nerve fibers running through the middle of the ratprostate (FIG. 16E). Higher magnification imaging showed that FAM-NP41additionally highlighted autonomic nerve branches surrounding theneurovascular bundle (FIG. 16G) which travel within the fatty capsule ofthe prostate gland. These branching nerves were not visible using whitelight reflectance imaging (FIG. 16F). To quantitate selective labelingof autonomic nerves in rats, nerves within the prostate gland wereimaged with both fluorescence and white light reflectance. Average nerveto non-nerve signal intensity from fluorescence was 1.275±0.02 (n=3)compared to 1.083±0.01 (n=3) for white light reflectance. To showapplicability to intra-surgical imaging we show that similar nervecontrast was observed in live rats using a clinical grade Zeiss Penteroimaging system (FIG. 16H). The Zeiss Pentero scope which is approved forclinical use overlays the fluorescent image from FAM-NP41 (yellow) onthe white light image with data collection in real time (FIG. 16H).Recordings during surgical manipulation show fluorescent fibers withinthe prostate that clearly present as nerves that are detectable usingNP41-FAM fluorescent guidance (data not shown). To confirm thatfluorescently labelled structures were indeed nerves, fluorescentsurgical guidance was used in real time to selectively dissect outfluorescent fibers that were thought to be nerves (FIG. 22: A).Dissected fluorescent fibers were then positioned vertically and flashfrozen in OCT embedding compound. Vertical cross sections were imagedusing fluorescence to show that suspected nerve fibers were centered onslides (FIG. 22: B). Fibers were confirmed to be nerve as they werefluorescently labelled using dual immunohistochemically analysis withantibodies against either fluorophore (FIG. 22: C) or tyrosinehydroxylase (FIG. 22: D), a known marker for unmyelinated autonomicnerves. No immunostaining was detected in the absence of primaryantibody (FIG. 22: E)

To enable translation of a nerve-illumination peptide for use in humanpatients, phage display was performed to identify human nerve bindingpeptides using an m13 phage library expressing 16 random amino acidsequences on the N-terminus of gill (Creative Biolabs). Phage wereselected using iterative rounds of selection for binding to human suralnerve with negative selection to muscle and fat. Counter selection tomuscles and fat was done by pre-absorbing library with these tissuesprior to selection for binding to human nerve. Individual phage weresequenced after each round of selection and three specific sequencesSGQVPWEEPYYVVKKS (HNP401; SEQ ID NO:1), and WEYHYVDLNWTSQHPQ (HNP402;SEQ ID NO:2) DLPDIIWDFNWETAG (HNP403; SEQ ID NO:3) were highly enrichedafter 5 and 6 rounds.

To test the affinity of selected phage display peptides for binding tohuman nerves, they were chemically synthesized by solid-phase synthesisand labeled with fluorescein at the C-terminus. Peptides were topicallyapplied to sections of surgically harvested human sural nerve andtemporalis muscle to determine nerve to muscle contrast for selectedpeptides and controls (FIG. 23). Controls including free dye(carboxyfluorescein) were also tested on various nerves from multiplepatient tissues to confirm specificity of peptide dye conjugates forbinding human nerve (FIG. 24). Free non-reactive dye control, such ascarboxyfluorescein, was shown to have only weak non-specific binding andare not efficacious for topical applications. FAM-HNP401 yielded thehighest contrast and was shown to be superior to the previously reportedrodent nerve binding peptide FAM-NP41 [22], when topically applied tohuman sural nerve (FIG. 17). To quantify differential binding to nerveversus muscle, fluorescence signal intensity was measured for ROIs fromthe perineurium of select nerves and human temporalis muscle. FAM-HNP401showed selective binding to human sural nerve with 10.9× fluorescentsignal intensity (1374.44±425.96) compared to FAM-NP41 (126.17±61.03)(FIG. 17D, p=0.009, Student's t-test, unpaired). Nerve to musclecontrast was comparable at 3.03±0.57 for FAM-HNP401 and 2.28±0.96 forFAM-NP41 (FIG. 17H, p=0.236, Student's t-test, unpaired).

FAM-HNP401 was also tested topically on ex-vivo tissue for labeling ofmouse facial nerve with surrounding muscle where it did not perform aswell as FAM-NP41 (FIG. 25: J-M). For comparison, ex-vivo tissue labelingof human laryngeal nerve with surrounding muscle with FAM-NP41 andFAM-HNP401 is shown (FIG. 25: F-I). Autofluorescence of human nervewithout treatment of peptide dye conjugate was negligible compared tosignal intensity acquired after topical application FAM-HNP401 (FIG.26). FAM-HNP401 also has a 2.3× higher signal intensity for in-vivobinding to mouse sciatic nerve compared to FAM-NP41 (FIG. 18 A-C). Nerveto surrounding muscle contrast is comparable for the two peptides (FIG.18D). FAM-HNP401 also highlighted rat sciatic nerve (FIG. 18E) andprostate nerve (FIG. 18F) at a dose of 2 μmole (^(˜)54 mg/kg) whenimaged 3 hours post injection. The bladder was drained with a syringeand sutured to avoid spillage and contamination around prostate. Thecollected urine was analyzed by mass spectrometry and as expectedfragments of the peptide with dye attached were detected indicatingpeptide in bladder was partially metabolized (FIG. 27). Autonomic nerveswithin the prostate and adjacent to the vascular bundle can be easilyvisualized when imaged at higher magnification using a dose of 0.5 μmole(13.4 mg/kg) FAM-HNP401 10 mins after probe injection (FIG. 18G). Bloodclearance of FAM-HNP401 showed a half-life of 30 minutes which issimilar to FAM-NP41 (FIG. 18H). Optimal nerve contrast was detectedusing 50-100 μM (FIGS. 25: A-E) with low concentration (10 μM) highresolution confocal imaging showing that FAM-HNP401 binds with higheraffinity to perineurium, epineurium and endoneurium while being excludedfrom axons (FIG. 25: N). FAM-HNP401 signal from human nerve saturates by100 μM while the signal from FAM-NP41 continues to increase even at 375μM but the signal intensity remains much lower than that of HNP401applied at the same concentration (FIGS. 25: F and H). Stability ofFAM-HNP401 in human plasma at 5 minutes and 2 hours was determined byincubation of peptide dye conjugate in human serum prior to analysis bymass spectrometry. For analysis the area under the curve at 450 nm andthe corresponding mass of FAM-HNP401 was determined after injection of afixed volume of analyte into the LC-MS (FIGS. 28:A-B). For comparison,we also tested the stability of FAM-NP41 in human plasma (FIGS. 28:C-D).Integration of the peak area at 5 minutes and 2 hours indicates thatboth FAM-HNP401 and FAM-NP41 were stable in human serum. Area ofextracted ion-current was used to determine peptide quantitation. Nodegradation of peptide-FAM conjugate was observed, with identicalconcentration detected at 5 min and 2 hours of incubation with humanplasma from analysis of the ion current. Peptides were analogouslytested and shown to be stable in rat cerebrospinal fluid following 2hour exposure (FIGS. 28:E-F).

FAM-HNP401 and FAM-NP41 were tested for binding to autonomic nerves,isolated from the prostate glands of two human patients (FIGS. 19 and29). FAM-HNP401 (FIGS. 19A and 29A) showed a significantly higherfluorescent signal in autonomic (cavernosal) nerves compared to FAM-NP41(FIGS. 19B and 29B). Quantitation was not done because only 2 patientsamples were available for testing as nerve resection during radicalprostatectomy is only performed in instances of gross capsular invasion.Labelled fibers were confirmed as nerve using anti-neurofilamentantibody SMI312 (red) with DAPI (blue) to show nuclear labeling (FIG.19C). H&E staining also confirmed label tissue as nerve by histology(FIG. 19D). SMI312 does not stain perineurium due to the lack ofneurofilament fibers in this region of the nerve bundle. SMI312 stainingshows that the tissue isolated is nerve due to staining of neurofilamentstructures that support the axons. Similar staining using FAM-HNP401 wasobtained for another sensory nerve (anti-brachial cutaneous) isolatedfrom human arm showing the broad nerve binding activity of HNP401 (FIGS.19E-H).

To optimize and attempt to determine the core binding domain of HNP401,systematic deletion of two amino acids from the C or N terminus wasperformed (see, for example, FIG. 30) followed by binding analysis onhuman sural nerve sections (FIG. 20). In each case nerve binding andsignal intensity was normalized to the parent FAM-HNP401 peptide (FIG.20J). Removal of the C-terminal serine (C-2) was tolerated but uponremoval of lysine (C-4) the solubility and binding was reduceddramatically with a normalized average signal intensity of 0.49±0.11 fornerve binding of HNP401-C-4 (FIG. 20F). Deletion of amino acids from Nterminal is mostly well tolerated. Removal of the N-terminal serine andglycine improved nerve selective binding about 2-fold with a normalizedaverage signal intensity of 2.02±0.65 for HNP401-N-2 (FIGS. 20A and 20J,p=0.026, Student's t-test, unpaired, one-tail). HNP401-N-4 has non-polaramino acids on its N-terminus which reduced binding to a normalizedaverage signal intensity of 0.56±0.18 (FIG. 20B). Removal of non-polaramino acids, tryptophan and proline, restored some binding intensityback to levels of FAM-HNP401 with HNP401-N-6 (FIG. 20C) and HNP401-N-8(FIG. 20D) having normalized average nerve signal intensities of1.0±0.34 and 0.98±0.31. The restored binding efficiency may be due toimproved solubility minimizing micro-aggregation that occurred when veryhydrophobic residues are present at the N terminus of peptide. C and Nterminal deletion studies of HNP-401 indicate the core binding domainlikely includes PYYVVKK with the N-terminal residues QVPWEE contributingto enhanced binding detected with HNP401-N-2. Normalized nerve totemporalis muscle contrast for HNP401-N-2 gave a 3-fold increase withrespect to FAM-HNP401 (FIG. 20K, p=0.011, Student's t-test, unpaired,one-tail).

Discussion

Various tracer substances have long been used to map the connectivity inthe nervous system although most of them have depended on anterograde orretrograde tracing after local application [11, 12, 26, 27]. Transportof tracers is relatively slow with contrast developing as dye moves awayfrom a the injection site [26, 27]. It is likely impractical to labelthe large areas exposed for surgeries by using these methods as multiplenerve tracts would have to be identified and independently labelled.There are reports of tracking retrograde neurovascular bundle and majorpelvic ganglion with lipophilic dyes in rodents [4, 28]. More recently,styryl pyridinium dyes [13-45], aminostyryl dyes[16-18], oxazine 4 [19,20], and anti-ganglioside antibodies [21] have been investigated invarious preclinical models to detect motor, sensory and autonomicnerves. Dyes alone have no selective mechanism for nerve targeting buttypically accumulate in the myelin. Myelin is known to be present in lowabundance or be absent in autonomic nerves which could limit the use offree dyes to highlight these fine but crucial nerves [29, 30]. Topicaland epidural application of free dyes has been used to locally labelnerves in animal models however these approaches may be limited inflexibility during human surgeries as tissue is removed and the field ofview changes [20, 31]. Anti-ganglioside antibodies have specifictargeting but have long blood half-lives which would likely requireinjection multiple days before surgery and may be more likely to elicitan immune response[32, 33]. Systemic injection of fluorescently labeledpeptides to label nerves overcomes the major disadvantages of thesetracers by labeling all nerves in the body with a single injection ofpeptide dye conjugate. We previously reported on NP41 for binding rodentmotor and sensory nerves and now demonstrate its potential applicationto the identification of fine autonomic nerves in rodent models. Wefound an average increase of 17% in nerve to non-nerve signal usingfluorescence imaging compared to contrast obtained by to white lightreflectance. This is a significant accomplishment given the unmyelinatednature of these nerves and their ultra-fine structure. However, topicalapplication of NP41 to human ex-vivo provided little contrast comparedto muscle. To enhance highlighting of human nerves with have nowidentified HNP401, a novel peptide that binds to and highlights humanmotor/sensory and autonomic nerves.

We expect the FAM-HNP-401 or optimized analog could enable clinicaltranslation of nerve visualization methods for use in surgeriesinvolving human patients. Fluorescently labeled HNP401 can bind andhighlight human sural, medial antebrachial cutaneous, laryngeal andautonomic nerves within and around the prostate gland. FAM-HNP401 showhigh signal intensity and reproducible labelling of nerve bundlescompared to its dye control of carboxyfluorescein. Carboxyfluoresceinshows low signal and non-specific binding to nerve on topical humannerve sections. Dyes such as FITC-isothiocyanate cannot be used as thecontrol as they will react with all nucleophilic side chains of proteinsexposed by cross-sectioning in unfixed tissue. Additionally,FITC-dextran, although clinical used, is not a viable control for ourexperiments as it labels vasculature including micro blood vessels deepwithin the nerve cross section and is a marker for nerve injury andneuropathic pain [34]. In addition, its large size affectspharmacokinetic profile of the dye. FAM-HNP401 consistently gave 10-foldhigher signal for binding human nerve compared to our previouslyidentified FAM-NP41 peptide dye conjugate. Higher signal intensity is anadvantage for real-time imaging requiring short exposure times. HNP401also showed a 3-fold contrast for nerve to muscle on topical sections inhuman ex-vivo tissue. FAM-HNP401 has a blood clearance profile similarto NP41 in mice [22]. FAM-HNP401 binds to myelinated and unmyelinatednerves. SMI312 antibody, which labels neurofilament does not colocalizedwith FAM-HNP401 staining demonstrating that FAM-HMP401 does not bindaxons, but preferentially binds the perineurium, and therefore may beless likely to affect nerve conductivity. It is this staining patternthat leads us to believe HNP401-FAM is binding structural protein(s) inthe perineurium. Polar amino acids at the C terminus appear to be neededfor both solubility and binding as removal either caused peptide tobecome significantly less soluble or show decreased binding affinity tonerves. Removal of 2 amino acids on the N terminus increased nervebinding but further deletions negatively affected both solubility andbinding. Attaching solubilizing groups like short PEGs may restorebinding to truncated variants.

For our initial studies nerve highlighting peptides HNP401 has beencoupled to fairly short wavelength fluorescein derivative to make itcompatible with dual nerve/tumor imaging with Cy5/Cy7 ratiometricactivatable cell penetrating peptides that are currently in phase IIclinical testing for detection of cancer (NCT03113825). Longerwavelength IR or near IR dyes such as indocyanine green (ICG), IRdye800would potentially allow nerves to be imaged deeper below the surface insurgically exposed tissue after attachment to HNP-401. Free oxazine 4has also been recently used to highlight nerves in preclinical modelsand targeting could be enhanced by coupling to targeting peptides likeHNP401. Although our preferred method of application is systemic,topical application is an option with some procedures. Such topicalapplication of dye to the exposed surface followed by a washing toremove unbound dye has been used to image nerve in animal models [20].Dyes such as 4-di-2-asp have also been used for topically application tonerves but it has the disadvantage of being toxic to nerves due itsbinding to mitochondria in nerve terminals [35]. Antibodies can beapplied intravenously or topically and have some advantages includinghigh affinity and a defined binding target, however as reported with theanti-ganglioside antibody they require long circulation times foraccumulation and washout to develop optimal nerve contrast.

In in-vivo rodent studies, we found that peripheral motor and sensorynerve can be labeled in mice at a dose of 150 nmoles FAM-NP41 whichwould easily scale to human dosing [36, 37]. Autonomic nerve labelingrequired a significantly higher dose in mice (600 nmols) so higheraffinity peptides like HNP401 or improved variants may be required foradvancement to clinical dosing. Interestingly, although higher dosingwas required to visualize very small autonomic nerve in rodents (assmall as 50 μm) labeling of significantly larger human prostate nerves(^(˜)750 μm) may be accomplished at a significantly reduced dose.Consistent with the conclusion that larger nerve can be highlighted witha lower dose, we were able to visualize nerve in rat prostate with a 40%dose NP41. Neither NP41 nor HNP401 permanently or covalently bind tonerve bundles as they both washes out with little remaining signal after24 hours. Structural proteins including laminins 421, 211 have beenidentified as the binding targets for NP41 [25]. While the bindingtargets for HNP401 is yet to be determined imaging data shows non-axonalbinding pattern similar to NP41 indicating it may also bind structuralnerve proteins. One significant characteristic of HNP401 compared tolipophilic dyes is that it does not require the presence of myelin andwe have shown that it can bind and highlight the neurovascular bundle aswell as the cavernosal nerve within the prostate. These nerves areimportant in urological applications and do not have high levels ofmyelination [29, 30]. We anticipate that preservation of nerves in thiscontext represent one of the most urgent unmet clinical needs [38] fornerve imaging technology. The ability of FAM-HNP401 to highlight thesenerves represents a significant advantage over competing nerve bindingagents that are selective for myelin [39] and incorporate into axons[21].

Methods Probe Synthesis

FAM-NP41 was synthesized as previously described [22]. A Prelude peptidesynthesizer and standard Fmoc solid phase peptide synthesis was used togenerate peptides with sequence acetyl-SGQVPWEEPYYVVKKSSGGC-CONH₂[HNP401], acetyl-WEYHYVDLNWTSQHPQGGC-CONH₂ [HNP402],acetyl-DLPDIIWDFNWETAGGC-CONH₂ [HNP403], each peptide having aC-terminal “GGC” linker. Carboxyfluorescein was conjugated to theC-terminal cysteine using 5-fluorescein-maleimide [Anaspec] in thepresence of N-methylmorpholine in DMSO. Peptides were purified onAgilent LCMS using a Phenomenex 5 um C18 Luna with mass and purity >95%confirmed by LC-MS. Truncated HNP401 peptides as listed in (see, forexample, FIG. 30) were synthesized and purified using the sameconfiguration and method described above.

Animals

Wild-type male SKH1 mice (Charles River, Wilmington, Mass.) weighing20-30 grams were used for testing of peptide dye conjugates. MaleSprague-Dawley rats weighing 100 to 250 grams were used for in-vivotesting of dye conjugates with dose being adjusted for based on animalsize. Protocols for use of animal were approved by the InstitutionalAnimal Care and Use Committee at University of California San Diego(Protocol number S05536).

In Vivo Imaging

Following anesthesia with intra-peritoneal injection of ketamine (80mg/kg) and midazolam (40 mg/kg), FAM-NP41 or its variants wereadministered into mice retro-orbitally. After a washout period of 2-4hours, the animals were anesthetized with ketamine (50 mg/ml) andxylazine (20 mg/ml). The bladder and prostate were exposed through amidline abdominal incision. The autonomic nerve along the cavernosalvessel in the prostate was imaged and recorded using a custom-madesurgical imaging system. This system is a modified from Olympus MVX10scope capable of hi-resolution fluorescence, RGB reflectance andrealtime overlay with zoom from 0.6 to 5.7 cm field of view. ImageJ wasused for quantitative analysis of nerve contrast for each peptide dyeconjugate tested. Images of autonomic nerve in prostate were selectedfrom the recorded files and magnified 300-400% prior to selection of ROIand measurement. Nerves and adjacent non-nerve tissues ROIs werehand-selected using polygonal selection tool at the same location fromboth of reflectance and fluorescence images. The mean and standarddeviation of the pixel intensities within the selected areas werecompared for nerves (mean=I_(n), SD=σ_(n)) and adjacent backgroundtissue (mean=I_(b), SD=σ_(b)). Nerve to non-nerve contrast wascalculated after background subtraction with formula|I_(n)−I_(b)|/(σ_(n) ²+σ_(m) ²)^(0.5). For imaging the nerves in theprostate gland of male rats peptide dye conjugate were injectedretro-orbitally. FAM-HNP401 was injected at a concentration of 13 mgs/kgfollowed by imaging after 15 minutes or alternatively a dose of 52mgs/kg was used with imaging after 3 hours. Live animal surgery wasperformed under a ketamine-xylazine cocktail according to IACUCprotocol. Sterile technique was used to expose the prostate; bladder wasdrained with a small syringe and sutured. The surgical field was washedwith sterile saline prior to imaging. Mann-Whitney test was used toanalyze data for both mice and rats to compare nerve intensity and nerveto non-nerve contrast between white light reflectance and fluorescence.

Confocal Imaging Parameters

Confocal data for FIG. 17 was acquired with 488 nm laser line, 10 μmsections on glass at 10× magnification, 0.45 NA air objective lens. Gainset to 50, power set to 0.5% of laser power, pixel dwell value of 1.2μs, aperture size of 1.2 μm and a pixel size of 0.26 with a 2k by 2ksize image. We used the Nyquist feature and acquired images as tiles toget maximum resolution.

Data for FIGS. 19A and 19B was acquired with 488 nm laser line, 10 μmsections on glass at 10× magnification, 0.45 NA air objective lens. Gainset to 40, power set to 3% of laser power, pixel dwell value of 1.2 μs,aperture size of 1.2 μm and a pixel size of 0.26 μm/px with a 2k by 2ksize image.

Data for FIGS. 19E and 19F was acquired with 488 nm laser line, 10 μmsections on glass at 10× magnification, 0.45 NA air objective lens. Gainset to 40, power set to 1% of laser power, pixel dwell value of 1.2 μs,aperture size of 1.2 μm and a pixel size of 0.3 pm/px with a 2k by 2ksize image.

SMI312 neurofilament antibody and Dapi staining were imaged at 10×magnification, 0.45 NA air objective lens, NA with gain of 50, power setto 5% of laser power for 405 nm laser line and gain of 100, power set to50% of laser power for 640 nm laser line. We used a pixel dwell of 3.2μs, aperture size of 1.2 μm and image size of 2k by 2k per tileresulting in a pixel size of 0.29 μm/px.

Data for FIG. 26 was acquired with 488 nm laser line, 10 μm sections onglass at 10× magnification, 0.45 NA air objective lens. Gain set to 40,power set to 3% of laser power, pixel dwell value of 2.4 μs, aperturesize of 1.2 μm and a pixel size of 0.3 μm/px with a 2k by 2k size image.

Dose response data set of FAM-HNP401 on human nerve tissue (FIGS.25:A-E) was acquired with 488 nm laser line, 10 μm sections on glass at10× magnification, 0.45 NA air objective lens. Gain set to 40, power setto 3% of laser power, pixel dwell value of 1.2 μs, aperture size of 1.1μm and a pixel size of 0.3 μm/px with a 2k by 2k size image.

Data for FIG. 29 was acquired with 488 nm laser line, 10 μm sections onglass at 25× magnification, 1.10 NA water immersion lens. Gain set to40, power set to 3% of laser power, pixel dwell value of 2.2 μs,aperture size of 1.2 μm and a pixel size of 0.11 pm/px with a 2k by 2ksize tiled image.

Phage Display

Phage display was done using a custom synthesized m13 phage libraries(diversity ^(˜)10⁹) expressing 16 random amino acid on the N-terminus ofgill (Creative Biolabs). The phage library was processed throughselections for binding to freshly resected or frozen human nerves assimilarly describe for the identification of NP41 which bound mousenerves [22]. Library was processed through up to 6 binding and washcycles. Prior to positive selection phage were counter-selected for highaffinity muscle and fat tissue binder by pre-absorbing library withthese tissues. For positive selection phage libraries were mixeddirectly with human sural nerve tissue and incubated for up to 2 hoursat 4° C. Following incubation, tissue phage mixtures were centrifugedand washed with PBS. Tissue pellets with bound phage were thenhomogenized, mixed with TG1 bacteria and plated on LB agar plates.Colonies were counted to determine titer followed by selecting singlecolonies for DNA preparation and sequencing. After each round ofselection phage were pooled and amplified for iterative selection. Phagethat were bound at each round were sequenced and repeats noted.Duplicate phage as shown in results were identified after 5 and 6 roundsof selection.

Topical Application on Tissue Sections and Imaging

Human sural nerve, antebrachial nerve and laryngeal nerve and temporalismuscle were obtained under IRB protocol number 130837 for Dr. QuyenNguyen. Human peripheral nerves (typically sural) were obtained frompatients undergoing nerve resection procedures. Human nerves fromprostate gland of two patients were acquired under Moores Cancer CentreBiorepository IRB protocol number 090401. Tissue were sectioned andmounted on glass slides or Cryojane tape. Tissue sections were placed ina humidifier chamber for 30 min before application of the peptidesolution. Peptides were diluted to appropriate concentration in 0.5×HBSSprior to topical application. 50 μl of peptide solution of with knownconcentration (1 μM to 375 μM) were applied to 10 μm nerve sections ontape or slides and incubated for 30 minutes in a humidifier chamber.After incubation with peptide nerve sections were washed with twice with0.5×HBSS and once with 1×PBS. A cover-slip was applied and slides wereimaged immediately on either a Zeiss Lumar dissecting scope of Nikon A1confocal microscope. For confocal imaging tissue sections of 10 μmthickness were imaged with 488 nm laser excitation 515 (25) and a 10×air objective and a 0.26 μm/pixel size. For immunohistochemistry theconfocal Images were acquired with a 20× air objective at 0.4 μm/pixel.

Image Analysis

Image J was used to analyze and compare images acquired using theconfocal microscope and the Lumar dissecting scope. For eachexperimental set where probes were compared, we kept the acquisitionparameters identical so as to directly compare the data obtained. Duringthe experiment, it is clear that FAM-HNP401 had the brightest signal inour topical application experiments. All raw image files for a givenexperimental cohort were loaded at the same time into Image J as 16-bittiff images. We then levelled the image for tissue treated withFAM-HNP401. Once these levels were set, the settings are propagated toall images in one step using Image J. The brightest image is set as thebenchmark for all other images in the cohort to avoid saturating whenthe leveling is propagated. For quantifying the images, regions ofinterest (ROI) were drawn and the signal counts measured in image J. ForFIG. 5, even though FAM-HNP401-N-2 is the brightest, for consistency wechoose FAM-HNP401 to level and normalize signal counts.

Immunofluorescence of Autonomic Nerves from Rat Prostate

Suspect unmyelinated nerve tissue was taken from prostate gland of malerat after in-vivo intravenous injection of TAMRA-NP41 (0.5 μmoles or11.3 mg/kg for 100 gm rat) visualized on custom-made surgicalfluorescence imaging system based on an Olympus dissecting microscope. 5Ξm cryosections of the tissue were generated using a Leica Cryostat andmounted on Cryojane tape. Tissue sections were fixed for 10 min with 4%para-formaldehyde in 1×PBS followed by a 1× PBS rinse. A 1:2000 dilutionof monoclonal antibody against TAMRA [Thermofisher Scientific Cat. No.MA1-041] (or polyclonal antibody against tyrosine hydroxylase [CellSignaling Technologies Prod. No. 2792S]) in 10% goat serum in PBS wereapplied; 20 μl per section and incubated overnight at room temperaturefollowed by a 1×PBS wash. A 1:500 dilution of biotinylated anti-mousesecondary antibody was applied in 10% goat serum in PBS to sections for2 hours followed by a 1×PBS wash. Vector RTU (avidin biotin complex) orAlexa 405 streptavidin was applied for 1 hour followed by a 1× PBS wash.Tissue was wet-mounted on slides with 1×PBS. Confocal Image was acquiredwith 20×air objective at resolution of 0.4 μm/pixel.

Immunofluorescence for Neurofilament

Fresh viable human nerve tissue was obtained from prostatectomy andfrozen in OCT blocks. 10 μm cryosections of tissue were mounted on glassTrue Bond slide. Hydrophobic barrier pen was applied to the glass aroundeach section. Tissue sections were fixed using 2% paraformaldehydeprepared in 1×PBS and washed 4 times with 1×PBS. 100 μl of blockingbuffer (0.01% Triton X solution, 1% BSA in 10% normal goat serum [Lifetechnologies 50062Z]) was applied for 30 min to each tissue section. Thetissue was then washed 4 times with 1×PBS and a 1:1000 dilution ofneurofilament antibody SMI312 antibody [Biolegend Cat. No. 837904] wasapplied to the tissue for overnight incubation at 4° C. Tissue waswashed 6 times with PBST. A 1:1000 dilution of anti-mouse secondaryantibody Alexafluor 555 was applied to the sections for 2 hours at 4° C.followed by washing with 1×PBS. Prolong Gold Anti-fade reagent with DAPI[Life Technologies P36931] was added prior to cover slipping andimaging.

H&E Staining Protocol

Tissue sections were fixed for 1 minute in 1:1 10% buffered formaldehydeand 200 proof ethanol. Slides were then washed with water and immersedin hematoxylin stain for 2 minutes. Slides were then washed withdistilled water and immersed in bluing solution for 30 seconds. Slideswere washed with distilled water and immersed in eosin solution for 1minute followed by wash with distilled water. Slides were sequentiallydipped in 50%, 95% and 100% ethanol to remove water. Slides were airdried and dipped in citrisolv before mounting a cover-slip withnon-xylene mounting solution and imaged on the Hamamatsu Nanozoomerusing bright-field at 20× magnification.

Blood Clearance for HNP401-FAM

Five 8-week-old SKH male mice were injected intravenously with 100 nmol[10.75 mg/kg for 25 gm mouse] of FAM-HNP401 in 100 μl of sterile water.Prior to blood draw mice were anesthetized with a 1:1 cocktail ofketamine: midazolam. Tail pricks were performed at 1 min, 10 min, 20min, 30 min, 1 hr, and 2 hrs after injection to collect 5 μl whole bloodwhich was dissolved in 100 μl Agilent ICP-MS tuning buffer. Samples werecentrifuged and equal volume of supernatants were analyzed using a Tecanfluorescence plate reader.

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The examples set forth above are provided to give those of ordinaryskill in the art a complete disclosure and description of how to makeand use the embodiments of the compositions, systems and methods of thedisclosure, and are not intended to limit the scope of what theinventors regard as their disclosure. Modifications of theabove-described modes for carrying out the disclosure that are obviousto persons of skill in the art are intended to be within the scope ofthe following claims. All patents and publications mentioned in thespecification are indicative of the levels of skill of those skilled inthe art to which the disclosure pertains. All references cited in thisdisclosure are incorporated by reference to the same extent as if eachreference had been incorporated by reference in its entiretyindividually.

All headings and section designations are used for clarity and referencepurposes only and are not to be considered limiting in any way. Forexample, those of skill in the art will appreciate the usefulness ofcombining various aspects from different headings and sections asappropriate according to the spirit and scope of the invention describedherein.

All references cited herein are hereby incorporated by reference hereinin their entireties and for all purposes to the same extent as if eachindividual publication or patent or patent application was specificallyand individually indicated to be incorporated by reference in itsentirety for all purposes.

Many modifications and variations of this application can be madewithout departing from its spirit and scope, as will be apparent tothose skilled in the art. The specific embodiments and examplesdescribed herein are offered by way of example only, and the applicationis to be limited only by the terms of the appended claims, along withthe full scope of equivalents to which the claims are entitled.

1. A method of identifying a human neuron or nerve comprising contactingthe human neuron or nerve with a targeting molecule comprising: apeptide sequence of SGQVPWEEPYYVVKKSS (HNP-401, SEQ ID NO:1) or avariant thereof having 2-8 amino acids deleted from the N-terminus orC-terminus; and a fluorescent moiety.
 2. The method of claim 1, whereinsaid targeting molecule comprises a peptide sequence ofSGQVPWEEPYYVVKKSS (HNP-401, SEQ ID NO:1); QVPWEEPYYVVKKSS (HNP401-N-2;SEQ ID NO:20); PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS(HNP401-N-6; SEQ ID NO:23); PYYVVKKSS (HNP401-N-8; SEQ ID NO:24);SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25); SGQVPWEEPYYVV (HNP401-C-4;SEQ ID NO:26); SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27); or SGQVPWEEP(HNP401-C-8; SEQ ID NO:28).
 3. The method of claim 1, wherein saidpeptide sequence further comprises a GC, GG, or GGC peptide sequence atthe C-terminus.
 4. The method of claim 3, wherein said peptide sequencehas an amino acid sequence selected from Ac-SGQVPWEEPYYVVKKSSGGC (HNP401with GGC linker; SEQ ID NO:4), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 withGGC linker; SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGClinker; SEQ ID NO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21),PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118),EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG(HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG(HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GGlinker; SEQ ID NO:123), and SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQID NO:124).
 5. The method of claim 1, wherein the fluorescent moiety isjoined to the N-terminus or C-terminus of the peptide.
 6. The method ofclaim 1, wherein the fluorescent moiety is joined to the peptide via alinker.
 7. The method of claim 6, wherein the linker is a straight orbranched-chain carbon linker, heterocyclic carbon linker, amino acidlinker, lipophilic residue, peptide linker, peptide nucleic acid linker,hydrazone linker, SPDB disulfide, sulfo-SPDB, maleimidomethylcyclohexane-1-carboxylate (MCC), aminohexanoic acid linker, polyetherlinker, or polyethylene glycol linker.
 8. The method of claim 1, whereinthe fluorescent moiety is selected from the group consisting of: afluorescent protein, a fluorescent peptide, a fluorescent dye, and anycombination thereof.
 9. The method of claim 1, wherein the fluorescentmoiety is selected from the group consisting of: a xanthene; a bimane; acoumarin; an aromatic amine; a benzofuran; a fluorescent cyanine; acarbazole; a dicyanomethylene pyrane; polymethine; oxabenzanthrane;pyrylium; carbostyl; perylene; acridone; quinacridone; rubrene;anthracene; coronene; phenanthrecene; pyrene; butadiene; stilbene;porphyrin; pthalocyanine; lanthanide metal chelate complexes; andrare-earth metal chelate complexes.
 10. The method of claim 1, whereinthe fluorescent moiety is selected from the group consisting of:5-carboxyfluorescein; fluorescein-5-isothiocyanate;6-carboxyfluorescein; tetramethylrhodamine-6-isothiocyanate;5-carboxytetramethylrhodamine; 5-carboxy rhodol; tetramethyl andtetraethyl rhodamine; diphenyldimethyl and diphenyldiethyl rhodamine;dinaphthyl rhodamine; rhodamine 101 sulfonyl chloride; Cy3, Cy3B, Cy3.5,Cy5, Cy5.5, Cy 7, indocyanine green, IR800CW, cyan fluorescent protein(CFP), EGFP, 6-FAM, FAM, fluorescein, 5,6-dicarboxyfluorescein, 5-(and6)-sulfofluorescein, sulfonefluorescein, succinyl fluorescein, 5-(and6)-carboxy SNARF-1, carboxyfluorescein sulfonate, carboxyfluoresceinzwitterion, carboxyfluorescein quaternary ammonium, carboxyfluoresceinphosphonate, carboxyfluorescein GABA, carboxyfluorescein-cys-Cy5,5′(6′)-carboxyfluorescein, fluorescein glutathione, and combinationsthereof.
 11. The method of claim 1, wherein the targeting moleculecomprises: Ac-SGQVPWEEPYYVVKKSSGGC-5FAM (HNP401 with GGC linker; SEQ IDNO:105), Ac-QVPWEEPYYVVKKSSGGC-5FAM (HNP401-N-2 with GGC linker; SEQ IDNO:108), Ac-PWEEPYYVVKKSSGGC-5FAM (HNP401-N-4 with GGC linker; SEQ IDNO:109), Ac-EEPYYVVKKSSGGC-5FAM (HNP401-N-6 with GGC linker; SEQ IDNO:110), Ac-PYYVVKKSSGGC-5FAM (HNP401-N-8 with GGC linker; SEQ IDNO:111), Ac-SGQVPWEEPYYVVKKGGC-5FAM (HNP401-C-2 with GGC linker; SEQ IDNO:112), Ac-SGQVPWEEPYYVVGGC-5FAM (HNP401-C-4 with GGC linker; SEQ IDNO:113), Ac-SGQVPWEEPYYGGC-5FAM (HNP401-C-6 with GGC linker; SEQ IDNO:114), Ac-SGQVPWEEPGGC-5FAM (HNP401-C-8 with GGC linker; SEQ IDNO:115), or 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQID NO:104).
 12. The method of claim 1, wherein said targeting moleculeis administered to a human subject.
 13. The method of claim 12, whereinsaid targeting molecule is administered intravenously or topically tothe human subject.
 14. The method of claim 12, wherein said targetingmolecule is administered prior to a surgical procedure on the subject.15. The method of claim 14, wherein said surgical procedure is performedon the head, neck, spine, heart, or prostate of the subject.
 16. Amethod of delivering a photosensitizing agent to a human neuron or nervecomprising contacting the human neuron or nerve with a targetingmolecule comprising: a peptide sequence of SGQVPWEEPYYVVKKSS (HNP-401,SEQ ID NO:1) or a variant thereof having 2-8 amino acids deleted fromthe N-terminus or C-terminus; and a photosensitizing agent.
 17. Themethod of claim 16, wherein the photosensitizing agent is selected fromthe group consisting of: a porphyrin, chlorin, and dye.
 18. The methodof claim 16, wherein the photosensitizing agent selected from the groupconsisting of: porphyrin, protoporfin IX, purlytin, verteporfin, HPPH,temoporfin, methylene blue, photofrin, protofrin, hematoporphyrin,Talaporfin, benzopophyrin monoacid, 5-aminileuvolinic acid, Lutetiumtexaphyrin, metallophthalocyanine,metallo-naphthocyaninesulfobenzo-porphyrazine,metallo-naphthalocyanines, zinc tetrasulfophthalocyanine,bacteriochlorins, metallochlorins, chlorine,Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein(DBF), IR700DX, naphthalocyanine, and porphyrin.
 19. The method of claim16, further comprising exposing the human neuron or nerve to a lightsource that activates the photosensitizing agent, wherein the humanneuron or nerve is ablated by the activated photosensitizing agent. 20.The method of claim 16, wherein said targeting molecule comprises apeptide sequence of SGQVPWEEPYYVVKKSS (HNP-401, SEQ ID NO:1);QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); PWEEPYYVVKKSS (HNP401-N-4;SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS(HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25);SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26); SGQVPWEEPYY (HNP401-C-6; SEQID NO:27); or SGQVPWEEP (HNP401-C-8; SEQ ID NO:28).
 21. The method ofclaim 16, wherein said peptide sequence further comprises a GC, GG, orGGC peptide sequence at the C-terminus.
 22. The method of claim 21,wherein said peptide sequence has an amino acid sequence selected fromAc-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4),Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8),Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9),Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12),Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13),Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21),PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118),EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG(HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG(HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GGlinker; SEQ ID NO:123), and SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQID NO:124).
 23. The method of claim 16, wherein the photosensitizingagent is joined to the N-terminus or C-terminus of the peptide.
 24. Themethod of claim 16, wherein the photosensitizing agent is joined to thepeptide via a linker.
 25. The method of claim 24, wherein the linker isa straight or branched-chain carbon linker, heterocyclic carbon linker,amino acid linker, lipophilic residue, peptide linker, peptide nucleicacid linker, hydrazone linker, SPDB disulfide, sulfo-SPDB,maleimidomethyl cyclohexane-1-carboxylate (MCC), aminohexanoic acidlinker, polyether linker, or polyethylene glycol linker.
 26. The methodof claim 16, wherein said targeting molecule is administered to a humansubject.
 27. The method of claim 26, wherein said targeting molecule isadministered intravenously or topically to the human subject.
 28. Themethod of claim 26, wherein said targeting molecule is administeredprior to a surgical procedure on the subject.
 29. The method of claim28, wherein said surgical procedure is performed on the head, neck,spine, heart, or prostate of the subject.